Effect of Cd⁺² on phosphate solubilizing abilities and hydrogen peroxide production of soil-borne micromycetes isolated from Phragmites australis-rhizosphere.

The aims of this work were to evaluate the phosphate-solubilization and hydrogen peroxide (H2O2) production by the soil-borne micromycetes, Aspergillus japonicus, Penicillium italicum and Penicillium dipodomyicola, isolated from Phragmites australis rhizosphere and to study the effect of several concentrations of Cadmium (Cd(2+)) on both variables. Our results showed that P. italicum achieved a higher P-solubilization and H2O2 production than A. japonicus and P. dipodomyicola, as only P. italicum showed a positive correlation (R(2) = 0.71) between P-solubilization and H2O2 production. In dose-response assays, P. italicum was also more tolerant to Cd(2+) (0.31 mM) in comparison to A. japonicus (0.26 mM). Analysis of the 2(4) factorial experimental design showed that P-solubilization by P. italicum was negatively affected by increases in Cd(2+) (p = 0.04) and yeast extract (p = 0.02) in the culture medium. The production of H2O2 was positively affected only by glucose (p = 0.002). Fungal biomass production was reduced significantly (p = 0.0009) by Cd(2+) and increased (p = 0.0003) by high glucose concentration in the culture medium. The tolerance and correlation between P-solubilization and H2O2 production in the presence of Cd(2+) was strain and species dependent. The effects of Cd(2+), glucose, ammonium sulfate and yeast extract on those variables were evaluated through a two-level factorial design. P. italicum is promising for P-solubilization in soils contaminated with Cd(2+) and may be an alternative for manufacture of biofertilizers to replace chemical fertilizers.

Role of plants in nitrogen and sulfur transformations in floating hydroponic root mats: A comparison of two helophytes.

Knowledge about the roles helophytes play in constructed wetlands (CWs) is limited, especially regarding their provision of organic rhizodeposits. Here, transformations of inorganic nitrogen and sulfur were monitored in a CW variety, floating hydroponic root mat (FHRM), treating synthetic wastewater containing low concentration of organic carbon. Two helophytes, Phragmites australis and Juncus effusus, were compared in duplicates. Striking differences were found between the FHRM of the two helophytes. Whereas ammonium was removed in all FHRMs to below detection level, total nitrogen of 1.15 ± 0.4 g m(-2) d(-1) was removed completely only in P. australis systems. The mats with J. effusus displayed effective nitrification but incomplete denitrification as 77% of the removed ammonium-nitrogen accumulated as nitrate. Furthermore, the P. australis treatment units showed on average 3 times higher sulfate-S removal rates (1.1 ± 0.45 g m(-2) d(-1)) than the systems planted with J. effusus (0.37 ± 0.29 g m(-2) d(-1)). Since the influent organic carbon was below the stoichiometric requirement for the observed N and S transformation processes, helophytes' organic rhizodeposits apparently contributed to these transformations, while P. australis provided about 6 times higher bioavailable organic rhizodeposits than J. effusus.

Transformation of chloroform in model treatment wetlands: from mass balance to microbial analysis.

Chloroform is one of the common disinfection byproducts, which is not susceptible to degradation and poses great health concern. In this study, the chloroform removal efficiencies and contributions of sorption, microbial degradation, plant uptake, and volatilization were evaluated in six model constructed wetlands (CWs). The highest chloroform removal efficiency was achieved in litter-added CWs (99%), followed by planted (46-54%) and unplanted CWs (39%). Mass balance study revealed that sorption (73.5-81.2%) and microbial degradation (17.6-26.2%) were the main chloroform removal processes in litter-added CWs, and that sorption (53.6-66.1%) and plant uptake (25.3-36.2%) were the primary contributors to chloroform removal in planted CWs. Around 60% of chloroform got accumulated in the roots after plant uptake, and both transpiration and gas-phase transport were expected to be the drivers for the plant uptake. Sulfate-reducing bacteria and methanogens were found to be the key microorganisms for chloroform biodegradation through cometabolic dechlorination, and positive correlations were observed between functional genes (dsrA, mcrA) and biodegradation rates. Overall, this study suggests that wetland is an efficient ecosystem for sustainable chloroform removal, and that plant and litter can enhance the removal performance through root uptake and microbial degradation stimulation, respectively.

Response of removal rates to various organic carbon and ammonium loads in laboratory-scale constructed wetlands treating artificial wastewater.

High levels (92 and 91%) of organic carbon were successfully removed from artificial wastewater by a laboratory-scale constructed wetland under inflow loads of 670 mg/m2 x d (100 mg/d) and 1600 mg/m2d (240 mg/d), respectively. Acidification to pH 3.0 was observed at the low organic carbon load, which further inhibited the denitrification process. An increase in carbon load, however, was associated with a significant elevation of pH to 6.0. In general, sulfate and nitrate reduction were relatively high, with mean levels of 87 and 90%, respectively. However, inhibition of nitrification was initiated with an increase in carbon loads. This effect was probably a result of competition for oxygen by heterotrophic bacteria and an inhibitory effect of sulfide (S2) toxicity (concentration approximately 3 mg/L). In addition, numbers of healthy stalks of Juncus effusus (common rush) decreased from 14 000 to 10 000/m2 with an increase of sulfide concentration, indicating the negative effect of sulfide toxicity on the wetland plants.

Performance evaluation using a three compartment mass balance for the removal of volatile organic compounds in pilot scale constructed wetlands.

To perform a general assessment of treatment efficiency, a mass balance study was undertaken for two types of constructed wetlands (CWs), planted gravel filters and plant root mat systems, for treating VOC (benzene; MTBE) polluted groundwater under field conditions. Contaminant fate was investigated in the respective water, plant, and atmosphere compartments by determining water and atmospheric contaminant loads and calculating contaminant plant uptake, thereby allowing for an extended efficiency assessment of CWs. Highest total VOC removal was achieved during summer, being pronounced for benzene compared to MTBE. According to the experimental results and the calculations generated by the balancing model, degradation in the rhizosphere and plant uptake accounted for the main benzene removal processes, of 76% and 13% for the gravel bed CW and 83% and 11% for the root mat system. Volatilization flux of benzene and MTBE was low (<5%) for the gravel bed CW, while in the root mat system direct contact of aqueous and gaseous phases favored total MTBE volatilization (24%). With this applied approach, we present detailed contaminant mass balances that allow for conclusive quantitative estimation of contaminant elimination and distribution processes (e.g., total, surface, and phytovolatilization, plant uptake, rhizodegradation) in CWs under field conditions.

Potential roles of anaerobic ammonium and methane oxidation in the nitrogen cycle of wetland ecosystems.

Anaerobic ammonium oxidation (anammox) and anaerobic methane oxidation (ANME coupled to denitrification) with nitrite as electron acceptor are two of the most recent discoveries in the microbial nitrogen cycle. Currently the anammox process has been relatively well investigated in a number of natural and man-made ecosystems, while ANME coupled to denitrification has only been observed in a limited number of freshwater ecosystems. The ubiquitous presence of anammox bacteria in marine ecosystems has changed our knowledge of the global nitrogen cycle. Up to 50% of N(2) production in marine sediments and oxygen-depleted zones may be attributed to anammox bacteria. However, there are only few indications of anammox in natural and constructed freshwater wetlands. In this paper, the potential role of anammox and denitrifying methanotrophic bacteria in natural and artificial wetlands is discussed in relation to global warming. The focus of the review is to explore and analyze if suitable environmental conditions exist for anammox and denitrifying methanotrophic bacteria in nitrogen-rich freshwater wetlands.

Batch methanogenic fermentation experiments of wastewater from a brown coal low-temperature coke plant.

Coke plant effluents with high contents of organic compounds are mainly treated by biological aerobic fermentation after physical pre-treatment. In this study, a brown coal condensate wastewater from a low temperature coking process was fermented under methanogenic conditions in discontinuous experiments. By this fermentation, acetate, propionate, and the main polyphenolic compounds (catechol, resorcinol and hydroquinone) were degraded to a level below the detection limit. The COD was reduced by 72% with a residual concentration of 2.1 g/L. This anaerobic fermented wastewater had a residual BOD5 of 0.66 g/L and 2.2 L CH4 were formed per litre of wastewater. An abiotic pre-treatment for this wastewater with air had a negative effect on the COD reduction and decrease of colour on the methanogenic fermentation due to the autoxidation of polyphenolic compounds to humic-like compounds. This study showed that methanogenic fermentations in the treatment sequence of brown coal coking wastewaters could reduce energy consumption for aeration in further treatment processes and had the potential for a better effluent quality due to a less formation of recalcitrant humic-like compounds.

Assessment of in situ biodegradation of monochlorobenzene in contaminated groundwater treated in a constructed wetland.

The degradation of monochlorobenzene (MCB) was assessed in a constructed wetland treating MCB contaminated groundwater using a detailed geochemical characterisation, stable isotope composition analysis and in situ microcosm experiments. A correlation between ferrous iron mobilisation, decreasing MCB concentration and enrichment in carbon isotope composition was visible at increasing distance from the inflow point, indicating biodegradation of MCB in the wetland. Additionally, in situ microcosm systems loaded with 13C-labelled MCB were deployed for the first time in sediments to investigate the biotransformation of MCB. Incorporation of 13C-labelled carbon derived from the MCB into bacterial fatty acids substantiated in situ degradation of MCB. The detection of 13C-labelled benzene indicated reductive dehalogenation of MCB. This integrated approach indicated the natural attenuation of the MCB in a wetland system. Further investigations are required to document and optimise the in situ biodegradation of MCB in constructed and natural wetland systems treating contaminated groundwater.

Effects of tidal operation on pilot-scale horizontal subsurface flow constructed wetland treating sulfate rich wastewater contaminated by chlorinated hydrocarbons.

Three different flow regimes were carried out in a pilot-scale horizontal subsurface flow constructed wetland-treating sulfate rich wastewater contaminated with monochlorobenzene (MCB) and perchloroethene (PCE). The three regimes were continuous flow, 7-day cycle discontinuous flow, and 2.5-day cycle discontinuous flow. The results show that intensifying the tidal regime (2.5-day cycle) significantly enhanced MCB removal before 2 m from the inlet and increasing PCE removal efficiency at 0.5 m. The PCE dechlorination process was promoted with tidal operation, especially under the 2.5-day cycle regime, with significant increases of cis-1,2- dichloroethenes (DCEs), vinyl chloride (VC), and ethene, but trans-1,2-DCE was significantly decreased after tidal operation. Due to the high sulfate concentration in the influent, sulfide was observed in pore water up to 20 and 23 mg L-1 under continuous flow and 7-day cycle regime, respectively. However, sulfide concentrations decreased to less than 4 mg L-1 under intensified tidal operation (2.5-day cycle). The increase of oxygen concentration in pore water through intensified tidal operation resulted in better MCB removal performance and the successful inhibition of sulfate reduction. In conclusion, intensifying tidal operation is an effective approach for the treatment of chlorinated hydrocarbons and inhibiting sulfide accumulation in horizontal subsurface flow constructed wetland.

The sulfur depot in the rhizosphere of a common wetland plant, Juncus effusus, can support long-term dynamics of inorganic sulfur transformations.

The sulfur cycle in the rhizosphere of constructed wetlands is frequently interlaced with transformations of carbon and nitrogen. Knowledge about the manifold sulfur transformations may thus aid in improving treatment performance of constructed wetlands. In this study, two laboratory-scale constructed wetland models (planted fixed bed reactors; PFR1 and PFR2) were used to investigate inorganic sulfur transformations at various total loads of sulfate and organic carbon. Sulfate, sulfide and elemental sulfur were the most abundant sulfur compounds detected, thus providing evidence for the simultaneous occurrence of dissimilatory sulfate reduction and sulfide oxidation. This co-occurrence was likely enabled by oxygen micro-gradients in the root-near environment, i.e. aerobic sulfide and elemental sulfur oxidation took place mostly at the roots while sulfate and elemental sulfur reduction occurred in the pore water under reduced redox conditions. The rhizosphere was found to be first sink, then source for sulfur during the course of the experiment. Immobilization of reduced sulfur was triggered by catabolism of organic matter coupled to dissimilatory sulfate reduction and the subsequent partial oxidation of generated sulfide. Good plant status was critical for sulfur deposition in the systems. Without externally provided sulfate the sulfur depot of the rhizosphere was a prolonged source for sulfur, which was remobilized into the pore water. Oscillations between sulfide and sulfur (PFR1) or sulfide and sulfate (PFR2) suggested a dynamic interplay between plants and various microbial guilds, i.e. dissimilatory sulfate and sulfur reducers on one side and sulfide and sulfur oxidizers on the other.

Removal of dimethylphenols and ammonium in laboratory-scale horizontal subsurface flow constructed wetlands.

Phenolic compounds in industrial wastewaters are toxic pollutants and pose a threat to public health and ecosystems. More recently, focus is being directed toward combining the treatment of these compounds with a cost-effective and environmentally sound technology. The removal efficiency of dimethylphenol and ammonium nitrogen was studied, for the first time, in three different laboratory-scale horizontal subsurface flow constructed wetlands planted with Juncus effusus. Two of the wetlands used were filled with gravel. One of these was planted and the second left without vegetation. The third wetland was a hydroponic system. It was found that the removal efficiencies of dimethylphenol was dependent on the inflow loading of the contaminant and was higher in the planted systems. Both planted systems yielded 99% removal efficiency up to loads of 240 mg/d, compared to only 73% for the unplanted constructed wetland. Factors and processes such as redox dynamics, methanogenesis, reduction of ammonium and low nitrate and nitrite concentrations imply simultaneous aerobic and anaerobic dimethylphenol transformations. A significant surplus of organic carbon was detected in the planted wetlands, which may originate from intermediates of the dimethylphenol transformation processes and/or organic plant root exudates. The present study demonstrates that horizontal subsurface flow constructed wetlands are a promising alternative system for the treatment of effluents contaminated with dimethylphenol isomers.

Sulfate removal and sulfur transformation in constructed wetlands: The roles of filling material and plant biomass.

Sulfate in effluent is a challenging issue for wastewater reuse around the world. In this study, sulfur (S) removal and transformation in five batch constructed wetlands (CWs) treating secondary effluent were investigated. The results showed that the presence of the plant cattail (Typha latifolia) had little effect on sulfate removal, while the carbon-rich litter it generated greatly improved sulfate removal, but with limited sulfide accumulation in the pore-water. After sulfate removal, most of the S was deposited with the valence states S (-II) and S (0) on the iron-rich gravel surface, and acid volatile sulfide was the main S sink in the litter-added CWs. High-throughput pyrosequencing revealed that sulfate-reducing bacteria (i.e. Desulfobacter) and sulfide-oxidizing bacteria (i.e. Thiobacillus) were dominant in the litter-added CWs, which led to a sustainable S cycle between sulfate and sulfide. Overall, this study suggests that recycling plant litter and iron-rich filling material in CWs gives an opportunity to utilize the S in the wastewater as both an electron acceptor for sulfate reduction and as an electron donor for nitrate reduction coupled with sulfide oxidation. This leads to the simultaneous removal of sulfate, nitrate, and organics without discharging toxic sulfide into the receiving water body.

Hydroponic root mats for wastewater treatment-a review.

Hydroponic root mats (HRMs) are ecotechnological wastewater treatment systems where aquatic vegetation forms buoyant filters by their dense interwoven roots and rhizomes, sometimes supported by rafts or other floating materials. A preferential hydraulic flow is created in the water zone between the plant root mat and the bottom of the treatment system. When the mat touches the bottom of the water body, such systems can also function as HRM filter; i.e. the hydraulic flow passes directly through the root zone. HRMs have been used for the treatment of various types of polluted water, including domestic wastewater; agricultural effluents; and polluted river, lake, stormwater and groundwater and even acid mine drainage. This article provides an overview on the concept of applying floating HRM and non-floating HRM filters for wastewater treatment. Exemplary performance data are presented, and the advantages and disadvantages of this technology are discussed in comparison to those of ponds, free-floating plant and soil-based constructed wetlands. Finally, suggestions are provided on the preferred scope of application of HRMs.

Effect of plants and filter materials on bacteria removal in pilot-scale constructed wetlands.

Due to the lack of testing units or appropriate experimental approaches, only little is known about the removal of bacteria in constructed wetlands. However, improved performance in terms of water sanitation requires a detailed understanding of the ongoing processes. Therefore, we analyzed the microbial diversity and the survival of Enterobacteriaceae in six pilot-scale constructed wetland systems treating domestic wastewater: two vertical sand filters, two vertical expanded clay filters and two horizontal sand filters (each planted and unplanted). Samples were taken from the in- and outflow, from the rhizosphere, and from the bulk soil at various depths. Colony-forming units of heterotrophic bacteria and coliforms were analyzed and the removal of bacteria between the in- and outflow was determined to within 1.5-2.5 orders of magnitude. To access the taxon-specific biodiversity of potential pathogens in the filters and to reduce the complexity of the analysis, specific primers for Enterobacteriaceae were developed. While performing PCR-SSCP analyses, a pronounced decrease in diversity from the inflow to the outflow of treated wastewater was observed. No differences were observed between the bulk soil of planted and unplanted vertical filters. Some bands appeared in the rhizosphere that were not present in the bulk soil, indicating the development of specific communities stimulated by the plants. The fingerprinting of the rhizosphere of plants grown on sand or expanded clay exhibited many differences, which show that different microbial communities exist depending on the soil type of the filters. The use of the taxon-specific primers enabled us to evaluate the fate of the Enterobacteriaceae entering the wetlands and to localize harboring in the rhizosphere. The most abundant bands of the profiles were sequenced: Pantoea agglomerans was found in nearly all samples from the soil but not in the effluent, whereas Citrobacter sp. could not be removed by the horizontal unplanted sand and vertical planted expanded clay filters. These results show that the community in wetland system is strongly influenced by the filtration process, the filter material and the plants.

Microbial nitrogen transformation in constructed wetlands treating contaminated groundwater.

Pathways of ammonium (NH4 (+)) removal were investigated using the stable isotope approach in constructed wetlands (CWs). We investigated and compared several types of CWs: planted horizontal subsurface flow (HSSF), unplanted HSSF, and floating plant root mat (FPRM), including spatial and seasonal variations. Plant presence was the key factor influencing efficiency of NH4 (+) removal in all CWs, what was illustrated by lower NH4 (+)-N removal by the unplanted HSSF CW in comparison with planted CWs. No statistically significant differences in NH4 (+) removal efficiencies between seasons were detected. Even though plant uptake accounted for 32-100 % of NH4 (+) removal during spring and summer in planted CWs, throughout the year, most of NH4 (+) was removed via simultaneous nitrification-denitrification, what was clearly shown by linear increase of δ(15)N-NH4 (+) with decrease of loads along the flow path and absence of nitrate (NO3 (-)) accumulation. Average yearly enrichment factor for nitrification was -7.9 ‰ for planted HSSF CW and -5.8 ‰ for FPRM. Lack of enrichment for δ(15)N-NO3 (-) implied that other processes, such as nitrification and mineralization were superimposed on denitrification and makes the stable isotope approach unsuitable for the estimation of denitrification in the systems obtaining NH4 (+) rich inflow water.

The dynamics of low-chlorinated benzenes in a pilot-scale constructed wetland and a hydroponic plant root mat treating sulfate-rich groundwater.

A rarely used hydroponic plant root mat filter (PRMF, of 6 m(2)) and a horizontal subsurface flow constructed wetland (HSSF CW, of 6 m(2)), operating in continuous flow and discontinuous outflow flushing modes, were investigated for treating sulfate-rich and organic carbon-lean groundwater contaminated with monochlorobenzene (MCB); 1,2-dichlorobenzene (1,2-DCB); 1,4-dichlorobenzene (1,4-DCB); and 2-chlorotoluene. Whereas the mean inflow loads ranged from 1 to 247 mg m(-2) days(-1), the range of mean inflow concentrations of the chlorobenzenes recorded over a period of 7 months was within 0.04 and 8 mg L(-1). A hydraulic surface loading rate of 30 L m(-2) days(-1) was obtained in both systems. The mean load removal efficiencies were found to vary between 87 and 93 % in the PRMF after a flow path of 4 m, while the removal efficiencies were found to range between 46 and 70 % and 71 to 73 % in the HSSF CW operating in a continuous flow mode and a discontinuous outflow flushing mode, respectively. Seasonal variations in the removal efficiencies were observed for all low-chlorinated hydrocarbons both in the PRMF and the HSSF CW, whereby the highest removal efficiencies were reached during the summer months. Sulfide formation occurred in the organic carbon-lean groundwater particularly in summer, which is probably due to the plant-derived organic carbon that fostered the microbial dissimilatory sulfate reduction. Higher redox potential in water was observed in the PRMF. In conclusion, the PRMF could be an option for the treatment of water contaminated with compounds which in particular need oxic conditions for their microbial degradation, such as in the case of low-chlorinated benzenes.

Treatment of industrial effluents in constructed wetlands: challenges, operational strategies and overall performance.

The application of constructed wetlands (CWs) has significantly expanded to treatment of various industrial effluents, but knowledge in this field is still insufficiently summarized. This review is accordingly necessary to better understand this state-of-the-art technology for further design development and new ideas. Full-scale cases of CWs for treating various industrial effluents are summarized, and challenges including high organic loading, salinity, extreme pH, and low biodegradability and color are evaluated. Even horizontal flow CWs are widely used because of their passive operation, tolerance to high organic loading, and decolorization capacity, free water surface flow CWs are effective for treating oil field/refinery and milking parlor/cheese making wastewater for settlement of total suspended solids, oil, and grease. Proper pretreatment, inflow dilutions through re-circulated effluent, pH adjustment, plant selection and intensifications in the wetland bed, such as aeration and bioaugmentation, are recommended according to the specific characteristics of industrial effluents.

Nitrogen transforming community in a horizontal subsurface-flow constructed wetland.

Constructed wetlands are important ecosystems with respect to nitrogen cycling. Here we studied the activity and abundance of nitrogen transforming bacteria as well as the spatial distribution of nitrification, anaerobic ammonium oxidation (anammox), and denitrification processes in a horizontal subsurface-flow constructed wetland. The functional genes of the nitrogen cycle were evenly distributed in a linear way along the flow path with prevalence at the superficial points. The same trend was observed for the nitrification and denitrification turnover rates using isotope labeling techniques. It was also shown that only short-term incubations should be used to measure denitrification turnover rates. Significant nitrate consumption under aerobic conditions diminishes nitrification rates and should therefore be taken into account when estimating nitrification turnover rates. This nitrate consumption was due to aerobic denitrification, the rate of which was comparable to that for anaerobic denitrification. Consequently, denitrification should not be considered as an exclusively anaerobic process. Phylogenetic analysis of hydrazine synthase (hzsA) gene clones indicated the presence of Brocadia and Kuenenia anammox species in the constructed wetland. Although anammox bacteria were detected by molecular methods, anammox activity could not be measured and hence this process appears to be of low importance in nitrogen transformations in these freshwater ecosystems.

Rhizosphere microflora of plants used for the phytoremediation of bitumen-contaminated soil.

The microbial communities and their degradative potential in rhizospheres of alfalfa (Medicago sativa) and reed (Phragmites australis) and in unplanted soil in response to bitumen contamination of soil were studied in pot experiments. According to the results of fluorescence microscopy, over a period of 27 months, bitumen contamination of soil reduced the total number of microorganisms more significantly (by 75%) in unplanted than in rhizosphere soil (by 42% and 7% for reed and alfalfa, respectively) and had various effects on some important physiological groups of microorganisms such as actinomycetes as well as nitrogen-fixing, nitrifying, denitrifying, ammonifying, phosphate-solubilizing, sulphur-oxidizing, cellulolytic and hydrocarbon-degrading microorganisms. The changes in the physiological structure of the microbial community under bitumen contamination were found to hinge on not merely the presence of plants but also their type. It was noted that the rhizosphere microflora of alfalfa was less inhibited by hydrocarbon pollution and had a higher degradative potential than the rhizosphere microflora of reed.

Development of constructed wetlands in performance intensifications for wastewater treatment: a nitrogen and organic matter targeted review.

The knowledge on the performance enhancement of nitrogen and organic matter in the expanded constructed wetlands (CWs) with various new designs, configurations, and technology combinations are still not sufficiently summarized. A comprehensive review is accordingly necessary for better understanding of this state-of-the-art-technology for optimum design and new ideas. Considering that the prevailing redox conditions in CWs have a strong effect on removal mechanisms and highly depend on wetland designs and operations, this paper reviews different operation strategies (recirculation, aeration, tidal operation, flow direction reciprocation, and earthworm integration), innovative designs, and configurations (circular-flow corridor wetlands, towery hybrid CWs, baffled subsurface CWs) for the intensifications of the performance. Some new combinations of CWs with technologies in other field for wastewater treatment, such as microbial fuel cell, are also discussed. To improve biofilm development, the selection and utilization of some specific substrates are summarized. Finally, we review the advances in electron donor supply to enhance low C/N wastewater treatment and in thermal insulation against low temperature to maintain CWs running in the cold areas. This paper aims to provide and inspire some new ideas in the development of intensified CWs mainly for the removal of nitrogen and organic matter. The stability and sustainability of these technologies should be further qualified.