Enhanced degradation of hydrocarbons in constructed wetlands aided with nutrients, surfactant, and aeration.

The use of constructed wetlands (CWs) is a promising approach for the remediation of hydrocarbon-polluted wastewater. The amendments of CWs with nutrients, surfactants, and aeration enhances the removal of pollutants from wastewater. The objective of the present study was to explore the effect of external stimulants, i.e., nutrients, surfactant, and aeration on hydrocarbons degradation potential of CWs. The CWs mesocosms were developed by the vegetation of Phragmites australis and amendments with nutrients (20 mg l-1 N, 2.6 mg l-1 P, and 16.4 mg l-1 K), surfactant Tween 20 (0.2%, v/v), and aeration (7 mg l-1) for the remediation of diesel-spiked water (2%, w/v). The comparative analysis showed that the addition of nutrients, surfactant, and aeration individually enhanced total petroleum hydrocarbons (TPHs) reduction, and maximum TPHs reduction (88.4%) was achieved after 60 days in the mesocosms amended with the combination of nutrients, surfactant, and aeration. Among different individual treatments, the aeration (alone) also played a pivotal role in TPHs reduction (61%). The least (12%) reduction in TPHs was achieved in the mesocosms supplied with surfactant only. This study revealed that the combined application of nutrients, surfactant, and aeration in CWs enhanced its hydrocarbons degradation performance.

The biodegradability of crude oil-polluted wastewater is not efficient. It is mainly due to the low bioavailability of hydrocarbons, and less amount of nutrients and dissolved oxygen in the wastewater. This study explores the importance of the amendments of nutrients, surfactant, and aeration on the enhanced performance of constructed wetlands (CWs) for the remediation of hydrocarbon-contaminated water. The application of nutrients, surfactant, and aeration in CWs not only enhanced hydrocarbon degradation and toxicity reduction but also improved plant growth.

Shade and salinity responses of two dominant coastal wetland grasses: implications for light competition at the transition zone.

BACKGROUND: Coastal wetlands are threatened by the increased salinity that may result from sea level rise. Salinity stress alters species zonation patterns through changes in competitive outcome between species differing in salinity tolerance. This study therefore aimed to understand how salinity and light affect two dominant and competing coastal wetland grasses that differ in salt tolerance, height and photosynthetic metabolism. METHODS: The C4 species Spartina anglica and the C3 species Phragmites australis were grown at five salinity levels (0, 7, 14, 21 and 28 ppt) and two light fluxes (100 % and 50 % of natural daylight) in an outdoor experimental setup for 102 d with full access to nutrients. KEY RESULTS: Salinity reduced the biomass, height and shoot density of P. australis from 81.7 g dry weight (DW), 0.73 m and 37 shoots per pot at a salinity of 0 ppt to 16.8 gDW, 0.3 m and 14 shoots per pot at a salinity of 28 ppt. Biomass, height and shoot density of S. anglica did not respond or were only slightly reduced at the highest salinity of 28 ppt. High salinity also resulted in a higher tissue concentration of N and P in P. australis. Both species had low ability to acclimate to the lower light flux. Shade acclimation in S. anglica occurred via modest changes in specific leaf area, pigment content and biomass allocation. CONCLUSIONS: High salinity reduced traits important for light competition and increased the nutrient concentration in P. australis leaf and root biomass, while this was overall unaffected in S. anglica. This is likely to reduce the competitive ability of P. australis over S. anglica for light because at high salinities the former cannot effectively shade the lower-growing S. anglica. Neither species effectively acclimates to shade, which could explain why S. anglica does not occur in the understorey of P. australis at low salinities.

Critical Review: Biogeochemical Networking of Iron in Constructed Wetlands for Wastewater Treatment.

Iron is present in all types of wastewater; however, besides acid mine drainage, where it is a major constituent of concern, it is usually neglected in other types of wastewaters. In all kinds of constructed wetlands, iron plays important role in removal of organics and phosphorus, and it has an impact on transformation of nitrogen, sulfur, and metals. The biogeochemistry of iron is well understood in natural wetlands, but knowledge about iron impact on microbiological and chemical transformations during wastewater treatment in constructed wetlands is very limited. So far, the sparse research in this area provides limited information on observed interactions with several varying parameters across the studies, making it difficult to draw fundamental and mechanistic conclusions. A critical review of the complex biogeochemical networking of iron in CWs is therefore necessary to fill the gap in knowledge on the role of iron and its biogeochemical multi-interactions in wastewater treatment processes of CWs. This review is the first with specific focus on iron, discussing its mitigation and retention in CWs with different configurations and operational strategies, and presenting both seasonal dynamics and the potential remobilization of Fe. It also comprehensively discusses the interactions of redox-controlled iron turnover with the biogeochemical processes of other elements, for example, carbon (C), nitrogen (N), phosphorus (P), sulfur (S), and heavy metals. The health response of wetland plants to both deficiency and toxicity of Fe in CWs designed with specific treatment targets has also been evaluated. Due to the complexity of various wastewater compositions and microredox gradients in the root rhizosphere in CWs, future research needs have also been identified.

Characterization of Hydrocarbon-Degrading Bacteria in Constructed Wetland Microcosms Used to Treat Crude Oil Polluted Water.

Ten plant species were grown in constructed wetlands (CWs) to remediate water containing 2% (w/v) crude oil. The plant species with better growth and biomass production were Typha latifolia and Cyperus laevigatus, and they were significantly correlated (R2 = 0.91) with hydrocarbon degradation. From T. latifolia and C. laevigatus, 33 hydrocarbon-degrading bacterial strains were isolated from the rhizosphere, and root and shoot interiors. More diversified bacteria were found in the rhizosphere and endosphere of C. laevigatus than those of T. latifolia. The predominant cultural hydrocarbon-degrading bacteria were shown to belong to the genera Pseudomonas, Acinetobacter and Bacillus. In addition to genes involved in hydrocarbon degradation, most of the bacteria displayed multiple plant growth promoting (PGP) activities. This study suggests the importance of selecting suitable bacterial strains with hydrocarbon degradation and PGP activities for improving the efficacy of CWs used in remediating water contaminated with crude oil.

Small genome separates native and invasive populations in an ecologically important cosmopolitan grass.

The literature suggests that small genomes promote invasion in plants, but little is known about the interaction of genome size with other traits or about the role of genome size during different phases of the invasion process. By intercontinental comparison of native and invasive populations of the common reed Phragmites australis, we revealed a distinct relationship between genome size and invasiveness at the intraspecific level. Monoploid genome size was the only significant variable that clearly separated the North American native plants from those of European origin. The mean Cx value (the amount of DNA in one chromosome set) for source European native populations was 0.490 ± 0.007 (mean ± SD), for North American invasive 0.506 ± 0.020, and for North American native 0.543 ± 0.021. Relative to native populations, the European populations that successfully invaded North America had a smaller genome that was associated with plant traits favoring invasiveness (long rhizomes, early emerging abundant shoots, resistance to aphid attack, and low C:N ratio). The knowledge that invasive populations within species can be identified based on genome size can be applied to screen potentially invasive populations of Phragmites in other parts of the world where they could grow in mixed stands with native plants, as well as to other plant species with intraspecific variation in invasion potential. Moreover, as small genomes are better equipped to respond to extreme environmental conditions such as drought, the mechanism reported here may represent an emerging driver for future invasions and range expansions.

Microbial density and diversity in constructed wetland systems and the relation to pollutant removal efficiency.

Microbes are believed to be at the core of the wastewater treatment processes in constructed wetlands (CWs). The aim of this study was to assess the microbial biomass carbon (MBC) and Shannon's diversity index (SDI) in the substrate of CWs planted with Phragmites australis, Hymenocallis littoralis, Canna indica and Cyperus flabelliformis, and to relate MBC and SDI to the pollutant removal in the systems. Significant higher MBC was observed in CWs with H. littoralis and C. indica than in CWs with P. australis, and the MBC differed with season and substrate depth. The microbial community in the wetlands included four phyla: Cyanobacteria, Proteobacteria, Chloroflexi, and Acidobacteria, with a more diverse community structure in wetlands with C. flabelliformis. The MBC in the substrate and the SDI of the 15-20 cm depth correlated with the removal of biochemical oxygen demand, NH4-N and NO3-N. Our results indicate that substrate SDI and MBC can both be regarded as bioindicators of the pollutant removal ability in CWs.

Increased invasive potential of non-native Phragmites australis: elevated CO2 and temperature alleviate salinity effects on photosynthesis and growth.

The prospective rise in atmospheric CO2 and temperature may change the distribution and invasive potential of a species; and intraspecific invasive lineages may respond differently to climate change. In this study, we simulated a future climate scenario with simultaneously elevated atmospheric CO2 and temperature, and investigated its interaction with soil salinity, to assess the effects of global change on the ecophysiology of two competing haplotypes of the wetland grass Phragmites australis, that are invasive in the coastal marshes of North America. The two haplotypes with the phenotypes 'EU-type' (Eurasian haplotype) and 'Delta-type' (Mediterranean haplotype), were grown at 0‰ and 20‰ soil salinity, and at ambient or elevated climatic conditions (700 ppm CO2, +5 °C) in a phytotron system. The aboveground growth of both phenotypes was highest at the elevated climatic conditions. Growth at 20‰ salinity resulted in declined aboveground growth, lower transpiration rates (E), stomata conductance (gs), specific leaf area, photosynthetic pigment concentrations, and a reduced photosynthetic performance. The negative effects of salinity were, however, significantly less severe at elevated CO2 and temperature than at the ambient climatic conditions. The Delta-type P. australis had higher shoot elongation rates than the EU-type P. australis, particularly at high salinity. The Delta-type also had higher maximum light-saturated rates of photosynthesis (Asat), maximum carboxylation rates of Rubisco (Vcmax), maximum electron transport rates (Jmax), triose phosphate utilization rates (Tp), stomata conductance (gs), as well as higher Rubisco carboxylation-limited, RuBP regeneration-limited and Tp-regeneration limited CO2 assimilation rates than the EU-type under all growth conditions. Our results suggest that the EU-type will not become dominant over the Delta-type, since the Delta-type has superior ecophysiological traits. However, the projected rise in atmospheric CO2 and temperature will alleviate the effects of salinity on both phenotypes and facilitate their expansion into more saline areas.

Ammonium tolerance and toxicity of Actinoscirpus grossus--a candidate species for use in tropical constructed wetland systems.

Actinoscirpus grossus, a native species in tropical wetlands of South-East Asia, North Australia and the Pacific islands, has been reported to perform well in experimental scale constructed wetland (CW) systems. However, little is known about how high NH4(+) concentrations prevailing in wastewater affect growth and performance of this species. We examined growth, morphological and physiological responses of A. grossus to NH4(+) concentrations of 0.5, 2.5, 5, 10 and 15mM under hydroponic growth conditions. The relative growth rates (RGR) of the plants were highest at 2.5mM NH4(+) but significantly reduced at 10 and 15mM NH4(+). The roots of the plants were stunted and produced subepidermal lignified-cell layers at exposure to 10 and 15mM NH4(+). The photosynthetic rates did not differ between treatments (average An=21.3±0.4µmolCO2m(-2)s(-1)) but the photosynthetic nitrogen and carbon use efficiency (PNUE and PCUE) were significantly depressed at 10 and 15mM NH4(+) treatments. The concentration of NH4(+) in the roots, but not in the leaves, reflected the NH4(+) concentration in the growth medium suggesting that the species is unable to regulate the NH4(+) uptake. The high root respiration rates in concert with high tissue NH4(+) and declined C/N ratio at 10 and 15mM NH4(+) suggest that the NH4(+) assimilation occurs primarily in the roots and the plant has inadequate C-skeletons for NH4(+) assimilation and exudation at high NH4(+) concentration in the external solution. The concentrations of mineral cations were generally reduced and the root membrane permeability increased at high external NH4(+) concentrations. Our study shows that A. grossus tolerates NH4(+) concentrations up to 5mM which is characteristic of most types of wastewater. Hence, A. grossus is a good native candidate species for use in CW systems in tropical and subtropical climates in South-East Asia, North Australia and the Pacific islands.

Emissions of CO2 and CH4 from sludge treatment reed beds depend on system management and sludge loading.

Sludge treatment reed beds (STRB) are considered as eco-friendly and sustainable alternatives to conventional sludge treatment methods, although little is known about greenhouse gas emissions from such systems. We measured CO2 and CH4 emissions and substrate characteristics in a STRB, an occasionally loaded sludge depot (SD) and a natural reed wetland (NW). The aim was to compare (i) emissions among the sites in relation to substrate characteristics and (ii) emissions before and after sludge loading in the STRB. The STRB emitted twice as much CO2 (1200 mg m(-2) h(-1)) as the SD, whereas the SD emitted four times more CH4 (2 mg m(-2) h(-1)) than the STRB. The NW had the lowest emissions of both gases. The differences in gas emissions among the sites were primarily explained by differences in the availability of oxygen in the substrate. As a consequence of overloading and poor management, the SD had no vegetation and a poor dewatering capacity, which resulted in anaerobic conditions favoring CH4 emission. In contrast, the well-managed STRB had more aerobic conditions in the sludge residue resulting in low CH4 emission rates. We conclude that well-designed and well-managed STRBs have a low climate impact relative to conventional treatment alternatives, but that overloading and poor sludge management enhances the emissions of CH4.

Chromosome-level genome assemblies reveal genome evolution of an invasive plant Phragmites australis.

Biological invasions pose a significant threat to ecosystems, disrupting local biodiversity and ecosystem functions. The genomic underpinnings of invasiveness, however, are still largely unknown, making it difficult to predict and manage invasive species effectively. The common reed (Phragmites australis) is a dominant grass species in wetland ecosystems and has become particularly invasive when transferred from Europe to North America. Here, we present a high-quality gap-free, telomere-to-telomere genome assembly of Phragmites australis consisting of 24 pseudochromosomes and a B chromosome. Fully phased subgenomes demonstrated considerable subgenome dominance and revealed the divergence of diploid progenitors approximately 30.9 million years ago. Comparative genomics using chromosome-level scaffolds for three other lineages and a previously published draft genome assembly of an invasive lineage revealed that gene family expansions in the form of tandem duplications may have contributed to the invasiveness of the lineage. This study sheds light on the genome evolution of Arundinoideae grasses and suggests that genetic drivers, such as gene family expansions and tandem duplications, may underly the processes of biological invasion in plants. These findings provide a crucial step toward understanding and managing the genetic basis of invasiveness in plant species.

Invasion of Old World Phragmites australis in the New World: precipitation and temperature patterns combined with human influences redesign the invasive niche.

After its introduction into North America, Euro-Asian Phragmites australis became an aggressive invasive wetland grass along the Atlantic coast of North America. Its distribution range has since expanded to the middle, south and southwest of North America, where invasive P. australis has replaced millions of hectares of native plants in inland and tidal wetlands. Another P. australis invasion from the Mediterranean region is simultaneously occurring in the Gulf region of the United States and some countries in South America. Here, we analysed the occurrence records of the two Old World invasive lineages of P. australis (Haplotype M and Med) in both their native and introduced ranges using environmental niche models (ENMs) to assess (i) whether a niche shift accompanied the invasions in the New World; (ii) the role of biologically relevant climatic variables and human influence in the process of invasion; and (iii) the current potential distribution of these two lineages. We detected local niche shifts along the East Coast of North America and the Gulf Coast of the United States for Haplotype M and around the Mississippi Delta and Florida of the United States for Med. The new niche of the introduced Haplotype M accounts for temperature fluctuations and increased precipitation. The introduced Med lineage has enlarged its original subtropical niche to the tropics-subtropics, invading regions with a high annual mean temperature (> ca. 10 °C) and high precipitation in the driest period. Human influence is an important factor for both niches. We suggest that an increase in precipitation in the 20th century, global warming and human-made habitats have shaped the invasive niches of the two lineages in the New World. However, as the invasions are ongoing and human and natural disturbances occur concomitantly, the future distribution ranges of the two lineages may diverge from the potential distribution ranges detected in this study.

Internal methane transport through Juncus effusus: experimental manipulation of morphological barriers to test above- and below-ground diffusion limitation.

Aerenchymatous plants can transport methane (CH(4) ) from the root zone to the atmosphere, bypassing the surface-oxidizing layers of the soil, yet morphological and anatomical factors that govern the transport of methane have rarely been critically tested in manipulative experiments. Here, we investigated the methane transport capacity of hydroponically grown Juncus effusus, in experiments with roots submerged in nutrient solutions sparged with methane (1.16 mmol CH(4) l(-1)). Through a range of manipulations of the above- and below-ground plant parts, we tested the contradictory claims in the literature regarding which sites provide the greatest resistance to gas transport. Root manipulations had the greatest effect on methane transport. Removing root material reduced methane transport significantly, and especially the lateral roots and the root tips were important. Cutting of the shoots, with or without subsequent sealing, did not alter methane transport significantly. We confirm modelling predictions that the limiting factor for methane transport in the tussock forming wetland graminoid, J. effusus, is the amount of permeable root surface, estimated using the proxy measurement of root length. The aerial tissues do not provide any significant resistance to methane transport, and the methane is emitted from the lower 50 mm of the shoots.

Tracing the origin of Gulf Coast Phragmites (Poaceae): a story of long-distance dispersal and hybridization.

PREMISE OF THE STUDY: Long-distance dispersal can affect speciation processes in two opposing ways. Dispersal can promote geographic isolation or it can bring together geographically distant and distantly related genotypes, thus counteracting local differentiation. We used the Gulf Coast of North America (GC), a "hot spot" of reed diversity and evolutionary dynamics, as a model system to study the diversification processes within the invasive, cosmopolitan, polyploid grass Phragmites. METHODS: Genetic diversity was studied using collections representing all species of the genus and from all continents (except Antarctica). A range of molecular markers, including chloroplast and nuclear sequences, microsatellites, and AFLPs, was analyzed to detect DNA variation from the population to the species level and to infer phylogenetic relationships across continents. KEY RESULTS: An interspecific hybrid, Phragmites mauritianus × P. australis, and four P. australis cp-DNA haplotypes from Africa, Europe, and North America have been dispersed to the GC and interbreed with each other. CONCLUSIONS: Long-distance dispersal and weak breeding barriers appear to be recurring phenomena, not only in the GC, but worldwide. We present data strongly suggesting that interspecific hybridization and introgression among different Phragmites species take place and appear to have contributed significantly to the diversification processes within the genus. Hence, the application of traditional species concepts within Phragmites might be inappropriate.

Nitrous oxide emission by aquatic macrofauna.

A large variety of aquatic animals was found to emit the potent greenhouse gas nitrous oxide when nitrate was present in the environment. The emission was ascribed to denitrification by ingested bacteria in the anoxic animal gut, and the exceptionally high N(2)O-to-N(2) production ratio suggested delayed induction of the last step of denitrification. Filter- and deposit-feeding animal species showed the highest rates of nitrous oxide emission and predators the lowest, probably reflecting the different amounts of denitrifying bacteria in the diet. We estimate that nitrous oxide emission by aquatic animals is quantitatively important in nitrate-rich aquatic environments like freshwater, coastal marine, and deep-sea ecosystems. The contribution of this source to overall nitrous oxide emission from aquatic environments might further increase because of the projected increase of nitrate availability in tropical regions and the numeric dominance of filter- and deposit-feeders in eutrophic ecosystems.

Response of Salvinia cucullata to high NH4(+) concentrations at laboratory scales.

Growth, morphology, NH(4)(+) uptake and mineral allocation in Salvinia cucullata Roxb. ex Bory grown with different amounts of NH(4)(+) were investigated. Plants of uniform size were grown on full strength Smart and Barko medium with different NH(4)(+) concentrations (0.5, 1, 5, 10 and 15 mM) and incubated in a greenhouse for four weeks. Salvinia cucullata grew well in the medium with 0.5-1 mM NH(4)(+) with a relative growth rate of 0.11-0.12 d(-1) without exhibiting NH(4)(+) toxicity symptoms. With an NH(4)(+) concentration above 5 mM, plant growth was suppressed and signs of NH(4)(+) toxicity were observed. NH(4)(+) toxicity symptoms were obvious in plants supplied with 10 mM and 15 mM NH(4)(+). These plants had low growth rates, short roots, low numbers of roots and showed chlorosis. Rotted roots and stems were also found in plants fed with 15 mM NH(4)(+). This species had a high uptake rate even though the NH(4)(+) concentrations increased, making it an ideal candidate for growth in eutrophic environments. The high NH(4)(+) concentration had a negative effect on K uptake resulting in low K concentration in the plant tissue, but the plants increased N content in plant tissue. Thus, harvested plants can be used as soil fertilizer or for animal feed. Furthermore, maintaining plant biomass can improve the efficiency of water treatment.

Sorption media for stormwater treatment--a laboratory evaluation of five low-cost media for their ability to remove metals and phosphorus from artificial stormwater.

Five sorption materials were studied with a focus on polishing pretreated stormwater: crushed limestone, shell-sand, zeolite, and two granulates of olivine. These materials are commercially available at comparatively low cost and have been subjected to a minimum of modification from their natural states. The sorbents were tested for phosphorus, arsenic, cadmium, chromium, copper, nickel, lead, and zinc at concentration and conditions relevant for typical stormwater. The materials were tested for sorption capacity and kinetics. Desorption was tested under neutral and alkaline conditions and in the presence of chloride. For most sorbent/sorbate combinations, significant sorption occurred within the first minutes of contact between sorbent and sorbate. Treatment to the low microgram per liter range could be achieved by contact times of less than 1 hour. The study indicated that sorption filters can be designed for long life expectancy at comparatively low cost by applying the materials tested.

Use of constructed wetland systems with Arundo and Sarcocornia for polishing high salinity tannery wastewater.

Treatment of tannery wastewater is problematic due to high and variable concentrations of complex pollutants often combined with high salinity levels. Two series of horizontal subsurface flow constructed wetlands (CWs) planted with Arundo donax and Sarcocornia fruticosa were set up after a conventional biological treatment system operating at a tannery site. The aim of the CWs was polishing organics and nitrogen from the high salinity effluent (2.2-6.6 g Cl(-) L(-1)). Both plant species established and grew well in the CW. Arundo, however, had more vigorous growth and a higher capacity to take up nutrients. The CWs were efficient in removing COD and BOD(5) with removal efficiencies varying between 51 and 80% for COD (inlet: 68-425 mg L(-1)) and between 53 and 90% for BOD(5) (inlet: 16-220 mg L(-1)). Mass removal rates were up to 615 kg COD ha(-1) d(-1) and 363 BOD(5) kg ha(-1) d(-1). Removal efficiencies were 40-93% for total P, 31-89% for NH(4)(+) and 41-90% for Total Kjeldahl Nitrogen. CW systems planted with salt tolerant plant species are a promising solution for polishing saline secondary effluent from the tannery industry to levels fulfilling the discharge standards.

Improved urban stormwater treatment and pollutant removal pathways in amended wet detention ponds.

Dissolved and colloidal bound pollutants are generally poorly removed from stormwater in wet detention ponds. These fractions are, however, the most bio-available, and therefore three wet detention ponds were amended with planted sand filters, sorption filters and addition of precipitation chemicals to enhance the removal of dissolved pollutants and pollutants associated with fine particles and colloids. The three systems treated runoff from industrial, residential and combined (residential and highway) catchments and had permanent volumes of 1,990, 6,900 and 2,680 m(3), respectively. The treatment performance of the ponds for elimination of total suspended solids (TSS), total nitrogen (Tot-N), total phosphorous (Tot-P), PO(4)-P, Pb, Zn, Cd, Ni, Cr, Cu, Hg were within the range typically reported for wet detention ponds, but the concentrations of most of the pollutants were efficiently reduced by the planted sand filters at the outlets. The sorption filters contributed to further decrease the concentration of PO(4)-P from 0.04 ± 0.05 to 0.01 ± 0.01 mg L(-1) and were also efficient in removing heavy metals. Dosing of iron sulphate to enrich the bottom sediment with iron and dosing of aluminium salts to the inlet water resulted in less growth of phytoplankton, but treatment performance was not significantly affected. Heavy metals (Pb, Zn, Cd, Ni, Cr and Cu) accumulated in the sediment of the ponds. The concentrations of Zn, Ni, Cu and Pb in the roots of the wetland plants were generally correlated to the concentrations in the sediments. Among 13 plant species investigated, Rumex hydrolapathum accumulated the highest concentrations of heavy metals in the roots (Concentration Factor (CF) of 4.5 and 5.9 for Zn and Ni, respectively) and Iris pseudacorus the lowest (CF < 1). The translocation of heavy metals from roots to the aboveground tissues of plants was low. Therefore the potential transfer of heavy metals from the metal-enriched sediment to the surrounding ecosystem via plant uptake and translocation is negligible.

Microbial Electrochemically Assisted Treatment Wetlands: Current Flow Density as a Performance Indicator in Real-Scale Systems in Mediterranean and Northern European Locations.

A METland is an innovative treatment wetland (TW) that relies on the stimulation of electroactive bacteria (EAB) to enhance the degradation of pollutants. The METland is designed in a short-circuit mode (in the absence of an external circuit) using an electroconductive bed capable of accepting electrons from the microbial metabolism of pollutants. Although METlands are proven to be highly efficient in removing organic pollutants, the study of in situ EAB activity in full-scale systems is a challenge due to the absence of a two-electrode configuration. For the first time, four independent full-scale METland systems were tested for the removal of organic pollutants and nutrients, establishing a correlation with the electroactive response generated by the presence of EAB. The removal efficiency of the systems was enhanced by plants and mixed oxic-anoxic conditions, with an average removal of 56 g of chemical oxygen demand (COD) mbed material -3 day-1 and 2 g of total nitrogen (TN) mbed material -3 day-1 for Ørby 2 (partially saturated system). The estimated electron current density (J) provides evidence of the presence of EAB and its relationship with the removal of organic matter. The tested METland systems reached the max. values of 188.14 mA m-2 (planted system; IMDEA 1), 223.84 mA m-2 (non-planted system; IMDEA 2), 125.96 mA m-2 (full saturated system; Ørby 1), and 123.01 mA m-2 (partially saturated system; Ørby 2). These electron flow values were remarkable for systems that were not designed for energy harvesting and unequivocally show how electrons circulate even in the absence of a two-electrode system. The relation between organic load rate (OLR) at the inlet and coulombic efficiency (CE; %) showed a decreasing trend, with values ranging from 8.8 to 53% (OLR from 2.0 to 16.4 g COD m-2 day-1) for IMDEA systems and from 0.8 to 2.5% (OLR from 41.9 to 45.6 g COD m-2 day-1) for Ørby systems. This pattern denotes that the treatment of complex mixtures such as real wastewater with high and variable OLR should not necessarily result in high CE values. METland technology was validated as an innovative and efficient solution for treating wastewater for decentralized locations.