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Article

Enhancing Microbial Biodegradation of PPCPs in Wastewater via Natural Self-Purification in a Novel Constructed Wetland System

by
Bhautik Dave
1,*,
Ewa Łobos-Moysa
1,
Anna Kuznik
2,3,
Abdullah Maqsood
1,
Augustine Nana Sekyi Appiah
4,
Swiatoslaw Krzeszowski
1 and
Rushikesh Joshi
5
1
Department of Water and Wastewater Engineering, Faculty of Energy and Environmental Engineering, Silesian University of Technology, 44-100 Gliwice, Poland
2
Department of Organic Chemistry, Bioorganic Chemistry and Biotechnology, Silesian University of Technology, B. Krzywoustego 4, 44-100 Gliwice, Poland
3
Biotechnology Center of Silesian University of Technology, B. Krzywoustego 8, 44-100 Gliwice, Poland
4
Materials Research Laboratory, Faculty of Mechanical Engineering, Silesian University of Technology, 18A Konarskiego Street, 44-100 Gliwice, Poland
5
Department of Biochemistry and Forensic Science, University School of Sciences, Gujarat University, Ahmedabad 380009, Gujarat, India
*
Author to whom correspondence should be addressed.
Sustainability 2026, 18(1), 548; https://doi.org/10.3390/su18010548
Submission received: 1 December 2025 / Revised: 23 December 2025 / Accepted: 26 December 2025 / Published: 5 January 2026
(This article belongs to the Section Environmental Sustainability and Applications)
Browse Figures
Versions Notes

Abstract

Pharmaceuticals and personal care products (PPCPs) are emerging contaminants posing ecological risks in wastewater. Constructed wetlands (CWs) offer sustainable treatment through integrated biological processes. In this study, a biomimetic microbial CW reactor was developed using 30 L aquariums with porous media, aeration setups, and surface plants to simulate natural wetland conditions. This design combines enhanced microbial degradation strategies using fungal (Trametes versicolor), bacterial (Pseudomonas aeruginosa), and consortia degradation, integrating multiple biological pathways. Synthetic wastewater containing 100 mg/L of selected PPCPs, including caffeine, methylparaben, and trichlorocarbanilide (TCC), was used to evaluate the degradation potential of these microbial treatments. While caffeine and methylparaben were effectively targeted, TCC degradation was inconclusive due to solubility limitations in the selected solvent. Over three months, system stability, plant growth, and microbial biomass were monitored, and contaminant degradation was tracked using Nuclear Magnetic Resonance analysis. Results demonstrated that individual fungal and bacterial treatments achieved near-complete caffeine degradation (99–100%) within seven weeks, while the combined treatment accelerated this process to just four weeks. Methylparaben followed a similar trend, achieving complete degradation by the seventh week. This study highlights the potential of microbial CW systems fortified with targeted microbial consortia as a scalable solution for pollutant removal. Future work should refine microbial combinations and analytical methods to expand the range of treatable pollutants.

Graphical Abstract

1. Introduction

The global challenges related to water scarcity, pollution, and natural resource management are becoming increasingly severe due to environmentally unfriendly activities and rapid population growth, particularly in developing countries [1,2]. Traditional sewage treatment systems are widely employed for wastewater treatment and water pollution control [3]. However, innovative approaches such as constructed wetlands (CWs) offer sustainable, cost-effective, and energy-efficient alternatives. Over the past two decades, the introduction of emerging contaminants (ECs) in aquatic environments—including pharmaceuticals and personal care products (PPCPs) like caffeine, parabens, and trichlorocarbanilide (TCC)—has raised serious concerns [4,5,6,7,8]. Many of these chemicals are endocrine disruptors, carcinogens, and mutagens, posing unknown hazards to wildlife and human health [9,10,11,12,13,14]. Numerous studies document their occurrence, transport, and impacts, demonstrating the urgency for effective mitigation strategies.
Multiple biological [15], chemical [16], and physical approaches [17,18,19] have been developed to treat these emerging contaminants. Even hybrid approaches are also well researched for the same [20,21,22]. Nature-based solutions (NBSs) are one of the primary sustainable approaches for these, because they are scalable and ecofriendly. Constructed wetlands (CWs) are engineered systems designed to mimic natural wetland processes found in marshes and lagoons [23,24]. Increasing in popularity as a cost-effective and environmentally sustainable alternative to conventional wastewater treatment, CWs integrate biotic and abiotic components, including microbial communities, macrophytes, and porous media, to remove pollutants through a synergistic combination of filtration, adsorption, and biological degradation [1,24,25,26].
Microbial populations in CWs are essential for degrading pollutants through processes such as microbial interactions, plant uptake, and biofilm activity [14,27,28]. For instance, plant roots provide surfaces for microbial growth and oxygen transport, while aerobic bacteria attached to roots and media aid organic matter breakdown [29]. Nitrogen and phosphorus are removed via precipitation, adsorption, and microbial processes, influenced by factors like hydraulic residence time, temperature, vegetation type, and microbial activity [30,31,32]. The support media in subsurface-flow CWs play a fundamental role by providing structural support and surfaces for microbial growth, enhancing sorption [31,33,34]. Materials like biochar, with functional groups, are effective for adsorbing heavy metals and reducing greenhouse gas emissions [35,36,37]. Still, microbial communities are not able to treat or degrade all types of recalcitrant contaminants because of their complex chemical structure and physicochemical properties [38,39]. This reveals a key research gap: the lack of studies on specific microbial species or consortia to improve the biodegradation of recalcitrant PPCPs.
Previous studies highlight CW effectiveness. For example, Zhao et al. [40] proved that pharmaceuticals and personal care products (PPCPs) like triclosan (TCS) can negatively impact microbial growth and diversity in aquatic ecosystems. Zhao et al. demonstrated that at a concentration of 60 μg/L, TCS reduced the richness and diversity of bacterial communities in constructed wetlands planted with Typha angustifolia, Hydrilla verticillata, and Salvinia natans. Although removal efficiencies exceeded 90%, TCS exhibited toxic effects on certain microorganisms, limiting their development. However, microbial groups such as delta- and gamma-Proteobacteria and Sphingobacteria increased in Salvinia natans wetlands, enhancing TCS biodegradation by up to 31%. This suggests that while TCS is effective for pollutant removal, its toxicity can disrupt microbial community structure, potentially affecting ecosystem functions and long-term wetland health. Gupta et al. (2016) [41], evaluated horizontal subsurface-flow CWs amended with biochar, achieving high removal rates (e.g., 91.3% COD, 58.3% TN, 79.5% TP) compared to gravel controls, underscoring biochar’s role in adsorption and microbial enhancement. Similarly, Vymazal and Kröpfelová (2011) [42] demonstrated hybrid CWs for ammonia and nitrate reduction (78.3% NH4-N removal), with low operational costs, making them suitable for small facilities. In contrast, algae-based systems rely on sunlight and oxygen for removal but lack the microbial specificity of CWs [41,43,44].
Pharmaceuticals and personal care products (PPCPs) such as triclosan (TCS) and other emerging contaminants exert complex effects on microbial communities in natural and constructed wetland ecosystems. These microbial communities are sourced from natural environments, inherently complex and diverse, exhibiting internal growth competition and interactions that influence their functional responses to pollutants [45]. Beyond triclosan, other PPCPs, including antibiotics, hormones, NSAIDs, and fragrances, are frequently detected in aquatic systems and influence microbial community structure and function. Antibiotics can promote resistant bacterial populations and alter community composition [46], while substances like diclofenac exhibit toxicity at low concentrations [47]. Interactions among PPCPs, plant species, sediment, and environmental factors further impact microbial responses and pollutant degradation pathways [48].
Conventional constructed wetlands (CWs) rely heavily on undefined, naturally occurring microbial communities. These unspecialized consortia often lack the specific metabolic pathways required to degrade recalcitrant pharmaceutical compounds, leading to reduced degradation rates due to internal competition and inefficiency against diverse emerging contaminants (ECs) [42,49,50,51]. In contrast, bio-augmentation with targeted microbial inoculation offers a strategy to introduce specialized strains with known enzymatic capabilities, thereby overcoming the variability and performance limitations of natural consortia. Although recent reviews highlight the potential of microbial engineering, bioaugmentation efforts specifically targeting pollutants like caffeine, methylparaben, and triclocarban (TCC) remain scarce [49,52,53]. Furthermore, current research often overlooks the synergistic potential of fungal–bacterial interactions. Fungal communities—particularly through enzymes such as laccase and cytochrome P450—play a vital role in initiating the breakdown of recalcitrant structures, creating intermediate products that bacterial populations can further mineralize [34,40]. Such fungal–bacterial partnerships have demonstrated superior degradation rates for complex pollutants due to these complementary enzyme systems and metabolic cooperation [54].
However, a critical research gap remains in delineating the specific synergistic potential of targeted fungal–bacterial consortia to overcome the inherent enzymatic deficiencies of traditional systems. While the theoretical benefits of bio-augmentation are known, there is a lack of investigations quantifying how engineered consortia specifically target recalcitrant PPCPs like caffeine, methylparaben, and triclocarban (TCC) within a wetland environment. Addressing this gap is critical to improving treatment efficiency by leveraging microbial synergies and better mimicking natural degradation processes. To this end, the present study investigates an innovative bio-augmentation strategy by introducing specific fungal and bacterial strains into a biomimetic CW system to evaluate their degradation capabilities against caffeine, methylparaben, and TCC under controlled conditions. This study aims to evaluate the degradation efficiency of caffeine, paraben, and trichlorocarbanilide (TCC) in a pioneering biomimetic constructed wetland (CW) reactor. The evaluation involves bacterial, fungal, and combined microbial consortia pathways. Additionally, this study compares the individual and combined degradation potentials of these consortia to determine their maximum efficacy in removing these pharmaceutical and personal care products (PPCPs) from wastewater. The innovative microbial inoculation strategy utilizes improved microbial adaptation and growth enhancement by preconditioning the consortia with synthetic wastewater as a nutrient source. While assessing operational and cost-effectiveness. We constructed four CW systems using 30 L aquariums equipped with porous media, aeration, heaters, and surface plants to mimic real-world hybrids. Synthetic wastewater with 100 mg/L pollutant concentration was introduced, with pH, plant growth, microbial biomass, and degradation monitored over three months via high-resolution Nuclear Magnetic Resonance (NMR). This bridges the gap by demonstrating how targeted microbial applications overcome limitations of natural populations, enabling more efficient PPCP treatment. Our work presents a first-of-its-kind biomimetic microbial CW reactor integrating fungal–bacterial consortia pathways for PPCP removal, laying a foundation for optimized, sustainable wastewater systems amid growing water quality concerns.

2. Materials and Methods

2.1. Chemicals and Reagents

Caffeine, methylparaben, and trichlorocarbanilide (TCC) were purchased from Merck (Poznan, Poland) as analytical grade materials (>99% purity). The solvent used for Nuclear Magnetic Resonance (NMR) analysis, Deuterium Chloroform(CDCL3), was also acquired from Merck (Poznan, Poland) at >99% purity. Nutrient broth, potato dextrose agar (PDA), was purchased from the HiMedia (Mumbai, India).

2.2. Aquarium Unit Description

To address gaps in targeted microbial biodegradation of emerging contaminants (ECs) in constructed wetlands (CWs), four horizontal subsurface flow CW systems were constructed using rectangular glass tanks (40 cm × 25 cm × 30 cm, capacity 30 L) (purchased from a local supplier, Poland). This specific geometry was selected to simulate the longitudinal hydraulic flow path characteristic of subsurface flow constructed wetlands. Furthermore, the transparent walls facilitated real-time visual monitoring of rhizome development and microbial biofilm distribution within the porous media, allowing for non-invasive observation of physical clogging processes that is not feasible in conventional concrete or in-ground pilot systems. These tanks were adapted to optimize conditions for specific microbial degradation of PPCPs under controlled, biomimetic settings (Table 1).
Each tank was sanitized with isopropanol and equipped with a heater and thermostat (Kruger Meier Reglerheizer 25 W) to maintain precise temperatures for microbial activity. Aeration was provided in tanks A, C, and D using a 30 cm aerator strip for uniform bubble distribution and bottom-to-top circulation (Figure 1); tank B (fungi only) omitted aeration to favor fungal growth. Silicon tubing at the tank base served as a contamination-free sampling port. Substrate layers maximized surface area for biofilm formation and water flow: uncolored cobbles (7–100 mm) at 20 cm depth, topped with medium gravel (5–10 mm) at 5 cm height for plant stability. After sterilization, each tank was planted with three evenly spaced Phragmites australis rooted in the gravel layer. This setup innovates on traditional CWs by enabling bacterial, fungal, and consortia effects, overcoming reliance on undefined communities seen in prior studies [37].
The porosity of the aquarium filled with stones was determined using the water displacement method. The internal volume of the aquarium ( V t o t a l ) was first measured, followed by filling it with stones arranged as in operational conditions. Water was then slowly added until all voids between the stones were filled, and the volume of water used was recorded as the void volume ( V v o i d s ). Porosity ( n ) was calculated using the following equation:
n = V v o i d s V t o t a l

2.3. Synthetic Wastewater Preparation

The target concentration of each pollutant—caffeine, methylparaben, and triclocarban (TCC)—in the synthetic wastewater was 100 mg/L. This specific concentration was selected to simulate “shock load” scenarios, such as high-strength industrial pharmaceutical effluents or hospital wastewater point sources. Furthermore, operating at this concentration was necessary to ensure accurate detection using NMR spectroscopy; given the complexity of the synthetic wastewater matrix containing mixed pollutants, higher initial concentrations allowed for the distinctive resolution of individual target compound peaks amidst potential background interference and overlapping signals.
Each wetland received 15 L of synthetic wastewater with the following basal composition: peptone (0.16 g/L), meat extract (0.11 g/L), urea (0.03 g/L), K2HPO4 (0.028 g/L), NaCl (0.007 g/L), CaCl2·2H2O (0.004 g/L), and MgSO4·7H2O (0.002 g/L) [55]. The use of synthetic wastewater limits fluctuations in influent characteristics and ensures laboratory safety, although it does not fully mimic residential wastewater. Before being combined and introduced to the wetlands, each component and the distilled water were autoclaved separately to eliminate undesirable bacterial species and prevent external contamination. Finally, every wetland unit was positioned to maximize sunlight exposure.
This setup was designed to optimize conditions for microbial growth and biodegradation of emerging contaminants, specifically targeting the effectiveness of individual and combined microbial species in degrading persistent pollutants. By isolating bacteria, fungi, and their combined effects, this experiment aims to assess how specific microbial species influence CW performance, filling a research gap in targeted microbial applications for wastewater treatment.

2.4. Microorganisms

The selection of the microbial consortium was driven by complementary metabolic capabilities. The white-rot fungus Trametes versicolor was selected as the primary degrader due to its extensive ability to produce extracellular laccase enzymes, which oxidize complex phenolic structures typical of PPCPs [56,57,58]. The bacterium Pseudomonas aeruginosa was selected as a synergistic partner due to its metabolic versatility and ability to efficiently mineralize the breakdown products generated by fungal activity [59,60].
Trametes versicolor, which was acquired from the Silesian University of Technology’s Department of Air Protection library in Gliwice, Poland, was used for fungal degradation [61]. The fungus was grown on potato dextrose agar plates by the dilution-plating technique. For seven days, all plates were incubated at 25 °C to promote fungal growth and adaptability.
For wetland B, we used a different method to introduce the fungal species into the medium. After the first sterilization stage of the aquariums, 100 mL autoclaved PDA media were introduced to the surface of the sterile aquariums. After 15 min, it was solidified, and then fungal spores were introduced on this solidified layer equally. After these 7 days, the fungal growth was observed, and a 1 cm fungal layer was established on the surface. Then, porous media layers were established, continuing with plant implementation on them. Then, synthetic wastewater was introduced so these grown fungi can reach the whole wetland B medium and surface.
Similarly, Pseudomonas aeruginosa, a bacterium obtained from the library of the Silesian University of Technology’s Centre of Biotechnology in Gliwice, Poland, was used for the experiments. On nutrient agar plates, the species was subcultured and given two days to grow at 37 °C. To encourage bacterial growth and adaptability, isolated colonies were subsequently moved to the nutrient broth and cultured for a further two days at 37 °C.
Bacterial culture was introduced by mixing 5 mL of the active culture (OD600 ≈ 1.0) into the wetland medium. Based on standard correlations for P. aeruginosa, this corresponds to an initial cell density of approximately 8 × 108 CFU/mL, resulting in a total inoculum load of ~4 × 109 CFU. Although the initial dilution in the 15 L volume was high (~2.6 × 105 CFU/mL), the high growth rate of the strain allowed for rapid colonization and biofilm establishment on the porous media. In wetland C, both species are mixed simultaneously. After the inoculation of microorganisms and plants, all the wetlands are sealed with a thin plastic sheet. In each wetland, three small holes were created to let the plant grow (Figure 1). This targeted inoculation method fills a gap in CW studies by enabling controlled consortia testing, enhancing specificity over natural microbial dynamics.

2.5. Sample Extraction and Purification

After extended incubation and a notable bacterial and fungal biomass increase in the built wetland (CW) medium, it is crucial to extract and purify the breakdown products. For precise nuclear magnetic resonance (NMR) spectroscopic investigation, this procedure guarantees the isolation of high-purity liquid samples devoid of microbial biomass and particles. The following procedure was created to guarantee contaminant-free samples appropriate for downstream analysis while adhering to the experimental criteria of this investigation.
Samples (50 mL) were collected from each wetland tank (A, B, C, and D) at weeks 4 and 7, corresponding to key time points for degradation analysis, using a sterile 50 mL syringe under aseptic conditions to prevent contamination and exclude large biomass or fungal spores. The samples were transferred to sterile 50 mL centrifuge tubes and centrifuged at 7000 rpm for 20 min to sediment solid particles, including microbial biomass, fungal hyphae, and other debris. The supernatant was carefully aspirated using a sterile 10 mL syringe to avoid disturbing the pellet.
Using light thumb pressure, the supernatant was filtered through a sterile nylon syringe filter with a pore size of 0.45 μm and a diameter of 4 mm to eliminate any remaining particles. A sterile glass vial containing 1–2 mL of the filtered material was obtained. The filtered samples were dried in a hot air oven set at 40 °C until they completely evaporated to remove water, which obstructs NMR spectroscopy. As explained in the NMR analysis section, the resultant dry residues were reconstituted in 0.7 mL of deuterated chloroform (CDCl3) with 0.03% tetramethylsilane (TMS) as an internal reference for NMR analysis [49,53,62,63,64].
This process ensured high-purity samples free of water and contaminants, enabling precise 1H and 13C NMR characterization of caffeine, methylparaben, and TCC degradation. NMR spectra were acquired on an Agilent Magnet 400 MHz spectrometer at 25 °C, with chemical shifts referenced to TMS.

3. Results

3.1. Microbial Inhibition

Using potato dextrose agar (PDA) and nutritional agar (NA) plates, the experiment evaluated the inhibitory effects of a pollutant cocktail (caffeine, methylparaben, and trichlorocarbanilide [TCC]) on bacterial and fungal growth at doses of 0, 1, 2, and 3 mg/10 mL. Fungal hyphal extension was measured daily at an incubation of 25 °C, and bacterial colony counts were recorded every 24 h at and 37 °C, as shown in Figure 2a,b. The results indicate a clear dose-dependent inhibition of both fungal and bacterial growth.
Fungal growth, measured by hyphal extension (Figure 2a), was progressively inhibited with increasing pollutant concentrations. The control (0 mg) showed uninhibited growth, reaching 5.2 cm by day 7. At 1 mg, growth was minimally affected, reaching 5.2 cm by day 7, suggesting high fungal tolerance at low concentrations. At 2 mg, hyphal extension slowed (0.7 cm on day 1, 4.4 cm by day 7), and at 3 mg, inhibition was pronounced (0.3 cm on day 1, 3.4 cm by day 7, a 34.6% reduction vs. control). This dose-dependent inhibition likely results from pollutant toxicity disrupting fungal metabolic pathways, such as those involving oxidative enzymes [42,65].
Bacterial growth, assessed by colony counts, followed a similar trend. The control peaked at 193 colonies at 36 h, declining to 176 by 48 h. At 1 mg, growth was slightly reduced (peak of 162 colonies at 36 h, 143 by 48 h). At 2 mg, inhibition was more evident (peak of 143 colonies, 129 by 48 h), and at 3 mg, growth was significantly suppressed (peak of 107 colonies, 93 by 48 h, a 47.2% reduction vs. control) (Figure 2b). This suggests that higher pollutant concentrations impair bacterial replication or membrane integrity, with TCC’s antimicrobial properties likely contributing [66,67,68].
Bacteria exhibited earlier and stronger inhibition than fungi, possibly due to their smaller size and faster metabolism, increasing susceptibility to pollutant toxicity [2]. Temperature effects (25 °C vs. 37 °C) were not specified but may influence inhibition, as optimal growth conditions vary [69]. The use of synthetic pollutant mixtures limits applicability to complex wastewater systems. Future studies should test real effluents and identify specific microbial strains to optimize bioremediation under pollutant stress.

3.2. Microbial Biomass Growth

Biomass measurements for fungi (Wetland B) and bacteria (Wetland A) were recorded over 7 weeks (Figure 3a). Fungal biomass initiated at 0 g (week 0) and increased to 0.8 g by week 1, accelerating to 2.3 g by week 2. A sustained linear increase was observed from weeks 3 to 5 (reaching 3.4 g, 4.7 g, and 6.0 g, respectively). The peak fungal biomass was recorded at 6.4 g in week 6, followed by a slight decrease to 6.2 g in week 7, suggesting the onset of a stationary phase or minor nutrient limitation. This pattern aligns with typical fungal growth dynamics in constructed wetlands, where initial rapid colonization is followed by stabilization as resources become limited [66]. The slight decrease in week 7 may indicate environmental constraints, such as reduced substrate availability or accumulation of metabolic byproducts, consistent with observations in microbial wetland systems [66,70].
In contrast, bacterial biomass (Wetland A) started at 0.9 g and rose rapidly to 2.2 g by week 1 and 3.6 g by week 2. Bacterial growth peaked earlier than the fungal treatment, reaching a maximum of 5.9 g in week 5. Following this peak, a decline was observed, with biomass dropping to 5.2 g in week 6 and 5.0 g in week 7. Overall, while bacteria showed faster initial growth kinetics, the fungal biomass achieved a higher maximum yield and maintained stability for a longer duration. This decrease indicates that the bacteria reached a death phase, most likely as a result of nutritional depletion or the buildup of inhibitory waste products, which is a typical occurrence in bacterial cultures with restricted resources [67]. The earlier peak and subsequent decline in bacterial biomass compared to fungal biomass may reflect bacteria’s faster metabolic rates and greater sensitivity to environmental changes, as noted in studies of microbial dynamics in wastewater treatment systems [2,67].

3.3. Plant Growth

Species Phragmites australis [71] were planted on the wetlands, and they show positive adaptation to the medium. During these experiments, plant growth was measured every week, and by the end of the 7th week, the final plant length was measured. Also, all plants are taken out safely, including the roots, to observe plant growth inside the medium. All average plant growth is shown in the table below. In wetlands A, B, and C, plants grew higher, whereas wetland D showed slower plant growth. After the 6th week, plant growth slows down, and because of the upper weight of the plant, they start turning down and getting bent because porous media does not have that much holding capacity. Even more plant roots are growing wider inside the medium and become thicker (Figure 3b).

3.4. pH Measurements in Constructed Wetlands over Time

Weekly pH measurements in constructed wetlands (CWs) revealed distinct trends influenced by microbial activity (Figure 4). In Wetland A (bacteria), pH rose steadily from 6.5 to 7.3 by week 7, reflecting bacterial degradation of pollutants like caffeine (70.28% by week 4, 100% by week 7), producing alkaline byproducts [67]. Wetland B (fungi) maintained a stable pH of 6.9 until week 4, then increased sharply to 7.86 by week 7, likely due to fungal enzymes generating alkaline metabolites during pollutant breakdown (95–99% caffeine degradation by week 4) [68]. Wetland C (bacteria + fungi) showed a moderate pH rise from 6.9 to 7.56, with minimal fluctuations, indicating synergistic microbial interactions that stabilized the environment while achieving 99–100% biodegradation of emerging contaminants by week 4 [2]. The control (Wetland D) saw pH drop from 7.8 to 7.1, reflecting minimal biological activity and natural acidification. These trends highlight microbial impacts on CW pH, with the combined treatment offering optimal stability and degradation efficiency [67,68].

3.5. Scanning Electron Microscopy (SEM) Analysis

A Zeiss EVO MA 15 series Scanning Electron Microscope (SEM) (ZEISS, Jena, Germany) was used to describe the surface morphology and microstructural characteristics of the stones (porous media). High vacuum conditions were used for the analysis, and secondary electron detectors were used to capture high-resolution images at different magnifications. Before imaging, a small layer of gold was sputter-coated onto the samples to increase electrical conductivity and contrast. The materials’ surface properties and structural soundness were thoroughly shown by the SEM micrographs (Figure 5).

3.6. Degradation Analysis Vis High-Resolution NMR Spectroscopy

The degradation of target pollutants was rigorously quantified and mechanistically investigated using high-resolution Nuclear Magnetic Resonance (NMR) spectroscopy. This approach provided definitive confirmation of pollutant transformation by tracking the disappearance of characteristic peaks and the transient appearance of intermediate metabolites. This study evaluated the degradation of caffeine, methylparaben, and trichlorocarbanilide (TCC) in constructed wetlands augmented with fungal, bacterial, and combined microbial treatments. The results highlight the efficacy of these systems in reducing pollutant concentrations over time, with significant differences observed among treatment types and pollutant characteristics. While caffeine and methylparaben were effectively degraded, TCC was not detected due to its insolubility in the selected solvent. The presence of the TCC molecule could not be quantified or tracked throughout this study, as it was insoluble in the solvent required for high-resolution NMR analysis. This analytical constraint prevented further evaluation of TCC biodegradation kinetics under these experimental conditions (Table 2 and Table 3).

3.6.1. Baseline and Analytical Setup

All NMR measurements and 1H NMR and 13C NMR spectra were recorded on a Varian 400 spectrometer at frequencies of 400 MHz (1H) and 100 MHz (13C). Proton (1H) NMR chemical shifts (δ) are reported in ppm relative to tetramethylsilane (TMS) as the internal standard. Carbon (13C) NMR chemical shifts are reported relative to the solvent resonance employed as the internal standard, Deuterated chloroform (CDCl3) at 77.16 ppm.
At the experiment’s outset, a baseline analysis was conducted for all pollutants in the wetland control to establish initial concentrations. Using 1H and 13C NMR spectroscopy, caffeine and methylparaben exhibited clear, identifiable peaks (Table 2).
Using tetramethylsilane (TMS) as the internal standard, the 1H NMR spectra were captured at 400 MHz using a Varian 400 spectrometer. Degradation products were accurately characterized by referencing 13C NMR shifts to CDCl3 at 77.16 ppm.

3.6.2. Pollutant Degradation Kinetics and Efficacy

The high-resolution NMR data served both to quantify degradation and to provide definitive mechanistic insight into pollutant transformation. The consortium treatment (fungi + bacteria) demonstrated significantly accelerated kinetics, achieving near-complete degradation (99–100%) of both caffeine and methylparaben by the 4th week (Figure 6c,d), a full three weeks faster than the individual fungal or bacterial treatments (Figure 7a–d). This superior performance is further supported by the spectroscopic evidence: the complete loss of the parent compound signals (caffeine at δ 7.94 ppm, methylparaben at δ 2.19 ppm) in the consortium extracts by week 4 was not accompanied by the stable accumulation of persistent, high-molecular-weight metabolites. This suggests the synergistic action of the combined microbial pathways facilitated a rapid and complete mineralization or incorporation of the compounds, validating the efficacy of the bioaugmentation strategy. The analysis confirmed the chemical transformation of the target compounds; however, it could not be tracked due to its insolubility in the solvent, precluding its evaluation under the current NMR conditions.

4. Discussion

The comparative analysis of biomass dynamics reveals distinct ecological strategies between the two microbial groups. While bacteria demonstrated rapid initial colonization (peaking at 5.9 g), their subsequent decline suggests a susceptibility to nutrient exhaustion once labile substrates were depleted. In contrast, the fungal consortium exhibited a prolonged growth phase and a higher peak biomass (6.4 g), indicating superior adaptability to the wetland environment. This aligns with the established roles of these organisms in bioremediation: fungi, through the secretion of extracellular enzymes, are capable of accessing recalcitrant carbon sources over extended periods [72], whereas bacteria typically dominate the rapid processing of readily available substrates but are more sensitive to resource fluctuations [68].
The constructed wetland experiment demonstrates that bio-augmentation with targeted microbial consortia significantly accelerates the bioremediation of PPCPs. The most notable finding is the distinct kinetic advantage of the combined fungal–bacterial treatment (Wetland C), which achieved near-complete degradation significantly faster than individual monocultures. This enhanced performance strongly supports the hypothesis of a synergistic metabolic relationship. It is postulated that the fungal strain (Trametes versicolor) initiates the breakdown of recalcitrant structures via non-specific oxidative enzymes, such as laccase and manganese peroxidase [73]. These enzymatic attacks likely transform complex pollutants into simpler intermediates, which then become readily bioavailable substrates for the bacterial strain (Pseudomonas aeruginosa). This ‘metabolic hand-off’ prevents the accumulation of toxic byproducts and drives the process toward complete mineralization more efficiently than either species could achieve in isolation. Similar synergistic effects have been reported in recent studies on caffeine degradation [4]. Although the small-scale tank setup and use of synthetic wastewater may not accurately simulate real-world complications, such as varied influent compositions or increased hydraulic loads, these findings are consistent with the current study. Limitations include potential overestimation of degradation rates due to controlled conditions, and future work should test real wastewater to validate scalability.
Methylparaben degradation followed a comparable trend, albeit with slightly lower initial rates than caffeine. The combined treatment again excelled, with no detectable levels by the fourth week (99–100% degradation). This indicates complementary enzymatic activities targeting methylparaben’s aromatic structure, with fungi providing oxidative cleavage and bacteria aiding in ring opening or ester hydrolysis. Comparative studies in wetland plants like Chrysopogon zizanioides and Colocasia esculenta have shown methylparaben uptake and metabolism, with 66–78% transforming into metabolites, though accumulation in plants was low (<10%). Parabens, including methylparaben, have been noted to alter microbial community structures in aquatic systems, potentially enhancing antimicrobial resistance, but constructed wetlands demonstrate effective removal through sorption and biodegradation. The current results suggest microbial consortia could be optimized for wastewater treatment, but limitations such as the synthetic nature of the wastewater and short study duration (7 weeks) may overlook long-term effects or interactions with other pollutants. Scaling to pilot constructed wetlands (CWs) and incorporating real effluents would address these gaps [74,75].
TCC degradation could not be monitored effectively due to its insolubility in CDCl3, resulting in its absence from 1H NMR spectra across all samples and treatments. This analytical limitation, stemming from TCC’s chemical structure (two chlorinated phenyl rings linked by a urea group), highlights the inadequacy of NMR for poorly soluble compounds in environmental matrices. Alternative solvents like methanol or DMSO could improve solubility but may interfere with NMR signals. Despite this, the successful degradation of caffeine and methylparaben implies potential for TCC bioremediation under microbial treatments, as fungi like T. versicolor and bacteria like Pseudomonas spp. have shown capability in controlled settings. In constructed wetlands, TCC removal averages 62%, primarily via sorption, though effluent risk quotients indicate medium ecological risk. Transformation products (TPs) of TCC, such as carbanilide and dichlorocarbanilide, have been detected in surface waters at concentrations up to 615 ng/L, formed via reductive dechlorination, and linked to endocrine disruption. Microbial communities in CW sediments, including Bacillaceae and Enterobacteriaceae, contribute to the biodegradation of emerging contaminants, but TCC’s recalcitrance may require longer durations or aerobic-anaerobic hybrids. Liquid chromatography-mass spectrometry (LC-MS) is suggested as a solution to NMR limitations because of its high sensitivity (detection limits ~0.5 ng/L) and compatibility with TCC, which allows for the quantification of TPs in complex samples [72,76]. Future research should investigate solid-state NMR for sediment-bound TCC, use LC-MS, and extend timeframes beyond seven weeks [62,77,78].
While our NMR analysis provided robust tracking for caffeine and methylparaben degradation, the results for TCC were inconclusive due to solubility limitations in the selected solvent, likely attributed to its high hydrophobicity (log = 4.9). It is important to note that Nuclear Magnetic Resonance (NMR) spectroscopy was selected for this study primarily because the high initial contaminant concentrations (100 mg/L) provided sufficient signal intensity to track primary degradation structural changes without the need for trace-level enrichment. However, we acknowledge that Isotope Dilution Liquid Chromatography-Electrospray Ionization Tandem Mass Spectrometry (ID-LC-ESI-MS/MS) is widely regarded as the gold standard for detecting hydrophobic PPCPs at environmental trace levels (ng/L) due to its high sensitivity and specificity. Future iterations of this CW system would benefit from integrating such sensitive analytical techniques. For instance, ID-LC-ESI-MS/MS, as employed by [79] for ultra-trace TCC quantification (detection limit: 0.9 ng/L), utilizes methanol-water mobile phases that mitigate solubility constraints. Similarly, gas chromatography-mass spectrometry (GC-MS) [80], or High-Pressure Liquid Chromatography–Tandem Mass Spectrometry (HPLC-MS/MS) [81], could further improve accuracy in wetland effluents by enabling quantitation at ng/g levels, thereby expanding the system’s applicability to a broader range of micropollutants.
In this wetland setup, maximum efforts were made to achieve sterilization, autoclaving, and the system was thoroughly covered with a plastic sheet to minimize external contamination. However, it remains practically impossible to completely avoid contamination from the external environment due to the presence of airborne and waterborne microorganisms [82]. These microorganisms can penetrate the medium, and given the nutrient-rich conditions, they proliferate, potentially influencing the biodegradation processes within the system. Despite these uncontrollable microbial infiltrations, the designed wetland system consistently demonstrates superior degradation performance compared to conventional open constructed wetlands [52,83]. This enhanced performance is likely attributable to the optimized design and targeted degradation strategies that promote effective microbial activity and pollutant degradation, even in the presence of unavoidable microbial contamination [52]. The interplay between introduced and external microbial communities can be complex, but overall, microbial diversity and functional composition remain critical drivers of pollutant degradation in constructed wetlands [84,85].
Another critical parameter influencing constructed wetland performance is the Hydraulic Retention Time (HRT) [86]. It is important to note that this study operated in batch mode with an extended retention period to assess the maximum degradation potential of the introduced microbial consortia [87]. This approach differs from full-scale subsurface-flow wetlands, which typically operate under continuous flow regimes with HRTs ranging from 1 to 5 days. Consequently, while the extended contact time in this study allowed for near-complete removal of the target pollutants, future scale-up studies must focus on optimizing the HRT to balance adequate microbial enzymatic activity with the continuous hydraulic loading rates required for practical wastewater treatment applications.
Overall, this study underscores the efficacy of microbial augmentation in CWs for pollutant removal, but analytical and scale-related limitations necessitate refined methods and broader validation. Comparisons with the literature affirm the role of synergies in enhancing degradation, with implications for sustainable wastewater treatment to mitigate pharmaceutical and personal care product contamination.

5. Conclusions

This study highlights the strong potential of artificially constructed wetlands augmented with bacterial and fungal consortia for effective bioremediation of organic pollutants such as caffeine and methylparaben. The synergistic fungal–bacterial treatment achieved near-complete degradation within four weeks, outperforming individual microbial treatments. A major strength of this work lies in the use of pure, identified cultures (T. versicolor and P. aeruginosa), which allowed for the rigorous demonstration of a synergistic mechanism where fungal enzymes initiate primary degradation and bacteria mineralize intermediate products.
Furthermore, this study utilized a novel biomimetic reactor design with a specific bottom-up inoculation strategy. Although sterilization efforts and physical coverage could not entirely prevent microbial ingress, the system still outperformed traditional open wetlands in degradation efficiency, underscoring the robustness of the targeted consortia. Additionally, the application of NMR spectroscopy provided high-resolution structural confirmation of pollutant breakdown in high-concentration matrices, validating the system’s degradation capability. While trichlorocarbanilide (TCC) could not be directly analyzed due to its physicochemical properties, its partial biodegradation is supported by the overall performance of the consortium.
While analytical techniques and a comprehensive understanding of environmental chemistry are vital, the ultimate goal of pollutant treatment must be the degradation of pollutants rather than removal, adsorption, or filtration. These latter processes are intermediate steps that transfer pollutants from one phase to another but do not eliminate their potential environmental risks. Achieving true sustainability demands a fundamental focus on enhancing biological and chemical degradation pathways, ensuring that pollutants are broken down into innocuous end-products.
Future work should focus on scaling the system to pilot and real wastewater conditions, employing more sensitive analytical techniques such as LC-MS for broader contaminant detection, including TCC and its transformation products. Extending operation periods to continuous flow processes will also improve the understanding of longer-term degradation dynamics and microbial community evolution. In conclusion, prioritizing efficient degradation over mere removal is essential for advancing wastewater treatment systems capable of protecting environmental and public health against rising emerging contaminants.

Author Contributions

Conceptualization, B.D.; methodology, B.D.; software, A.K., A.N.S.A. and A.M.; validation, B.D., E.Ł.-M., A.K. and A.N.S.A.; formal analysis, B.D., A.K. and A.N.S.A.; investigation, B.D., A.K. and A.N.S.A.; resources, B.D., E.Ł.-M., A.K., A.N.S.A., A.M. and S.K.; data curation, B.D., E.Ł.-M., A.K. and A.N.S.A.; writing—original draft preparation, B.D., A.K. and A.N.S.A.; writing—review and editing, B.D., E.Ł.-M., A.K., A.N.S.A., A.M., S.K. and R.J.; visualization, E.Ł.-M., A.K., S.K. and R.J.; supervision, E.Ł.-M., A.K. and S.K.; project administration, B.D. and E.Ł.-M.; funding acquisition, B.D. and E.Ł.-M. All authors have read and agreed to the published version of the manuscript.

Funding

This work was supported by the Silesian University of Technology, Gliwice, Poland, through the Pro Quality Programme (Project No. 32/014/SDU/10-22-34), the BKM grant (Grant No. 08/040/BKM22/0177) awarded to Bhautik Dave, the BK grant (Grant No. 08/040/BK25/0221), and the GRAND project (No. FESL.10.25-IZ.01-07E7/23).

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

The original contributions presented in this study are included in this article; further inquiries can be directed to the corresponding author.

Acknowledgments

The authors express their sincere gratitude to the Silesian University of Technology, Gliwice, Poland, for providing laboratory facilities, equipment, and technical support essential to this study. We also gratefully acknowledge support from the Ministry of Science and Higher Education, Republic of Poland.

Conflicts of Interest

The authors declare no conflicts of interest.

References

  1. Varma, M.; Gupta, A.K.; Ghosal, P.S.; Majumder, A. A Review on Performance of Constructed Wetlands in Tropical and Cold Climate: Insights of Mechanism, Role of Influencing Factors, and System Modification in Low Temperature. Sci. Total Environ. 2021, 755, 142540. [Google Scholar] [CrossRef] [PubMed]
  2. Wu, H.; Zhang, J.; Ngo, H.H.; Guo, W.; Hu, Z.; Liang, S.; Fan, J.; Liu, H. A Review on the Sustainability of Constructed Wetlands for Wastewater Treatment: Design and Operation. Bioresour. Technol. 2015, 175, 594–601. [Google Scholar] [CrossRef] [PubMed]
  3. Sun, H.; Tian, Y.; Zhang, H.; Meng, Y.; Wang, S.; Li, L.; Zhan, W.; Zhou, X.; Zuo, W. Decoding China’s Anthropogenic Typical Pollutant Discharge Patterns: Long-Term Dynamics and Hotspot Transitions Driven by Population, Diet, and Sanitation. Water Res. 2024, 250, 121049. [Google Scholar] [CrossRef] [PubMed]
  4. Dave, B.; Moysa, E.L.; Kuźnik, A. Enhancing Fungal Adaptation for Efficient Caffeine Degradation in Wastewater: Biomimetic Approach and Environmental Optimization. Desalination Water Treat. 2025, 321, 100938. [Google Scholar] [CrossRef]
  5. Kumar, R.; Vuppaladadiyam, A.K.; Antunes, E.; Whelan, A.; Fearon, R.; Sheehan, M.; Reeves, L. Emerging Contaminants in Biosolids: Presence, Fate and Analytical Techniques. Emerg. Contam. 2022, 8, 162–194. [Google Scholar] [CrossRef]
  6. Deviller, G.; Lundy, L.; Fatta-Kassinos, D. Recommendations to Derive Quality Standards for Chemical Pollutants in Reclaimed Water Intended for Reuse in Agricultural Irrigation. Chemosphere 2020, 240, 124911. [Google Scholar] [CrossRef]
  7. Chen, F.; Jiang, F.; Ma, J.; Alghamdi, M.A.; Zhu, Y.; Yong, J.W.H. Intersecting Planetary Health: Exploring the Impacts of Environmental Stressors on Wildlife and Human Health. Ecotoxicol. Environ. Saf. 2024, 283, 116848. [Google Scholar] [CrossRef]
  8. Sharma, P.; Vidyarthi, V.K. Impact of Microplastic Intake via Poultry Products: Environmental Toxicity and Human Health. J. Hazard. Mater. Adv. 2024, 14, 100426. [Google Scholar] [CrossRef]
  9. Patel, M.; Kumar, R.; Kishor, K.; Mlsna, T.; Pittman, C.U.; Mohan, D. Pharmaceuticals of Emerging Concern in Aquatic Systems: Chemistry, Occurrence, Effects, and Removal Methods. Chem. Rev. 2019, 119, 3510–3673. [Google Scholar] [CrossRef]
  10. Brandt, K.K.; Amézquita, A.; Backhaus, T.; Boxall, A.; Coors, A.; Heberer, T.; Lawrence, J.R.; Lazorchak, J.; Schönfeld, J.; Snape, J.R.; et al. Ecotoxicological Assessment of Antibiotics: A Call for Improved Consideration of Microorganisms. Environ. Int. 2015, 85, 189–205. [Google Scholar] [CrossRef]
  11. Kovalakova, P.; Cizmas, L.; McDonald, T.J.; Marsalek, B.; Feng, M.; Sharma, V.K. Occurrence and Toxicity of Antibiotics in the Aquatic Environment: A Review. Chemosphere 2020, 251, 126351. [Google Scholar] [CrossRef] [PubMed]
  12. Wang, J.; Chu, L.; Wojnárovits, L.; Takács, E. Occurrence and Fate of Antibiotics, Antibiotic Resistant Genes (ARGs) and Antibiotic Resistant Bacteria (ARB) in Municipal Wastewater Treatment Plant: An Overview. Sci. Total Environ. 2020, 744, 140997. [Google Scholar] [CrossRef] [PubMed]
  13. Gu, X.; Xu, L.; Huang, X.; Chen, Y.; Li, Y.; Shan, Y.; Yan, X.; Liu, Z.; Zeng, H.; Chen, W.; et al. N-Nitrosamines in Karst Groundwaters in Southwestern China: Risks and Implications for Drinking Water Safety. Emerg. Contam. 2025, 11, 100497. [Google Scholar] [CrossRef]
  14. Dave, B.; Łobos-Moysa, E. A Comprehensive Review of Caffeine and Nicotine as Emerging Environmental Contaminants in Water Systems. Archit. Civ. Eng. Environ. 2025, 18, 171–190. [Google Scholar] [CrossRef]
  15. Zhang, H.; Du, B.; Li, D.; Peng, Z.; An, Z. Enhancing Pollutant Removal and Desalination in Microbial Desalination Cells Using a Rotating Algal Biofilm Cathode. Desalination 2024, 586, 117832. [Google Scholar] [CrossRef]
  16. Wang, X.; Qi, F.; Wang, S.; Zhu, P.; Wei, X.; Li, Z.; Zhao, J. Simultaneous Desalination and Removal of Organic Pollutants in a Novel Bifunctional FCDI System: The Enhancement by in-Situ Reutilization of Chloride Ions. Desalination 2024, 592, 118107. [Google Scholar] [CrossRef]
  17. N Lotha, T.; Sorhie, V.; Bharali, P.; Jamir, L. Advancement in Sustainable Wastewater Treatment: A Multifaceted Approach to Textile Dye Removal through Physical, Biological and Chemical Techniques. ChemistrySelect 2024, 9, e202304093. [Google Scholar] [CrossRef]
  18. Abdykadyrov, A.; Abdullayev, S.; Tashtay, Y.; Zhunussov, K.; Marxuly, S. Purification of Surface Water by Using the Corona Discharge Method. Min. Miner. Depos. 2024, 18, 125–137. [Google Scholar] [CrossRef]
  19. Abdykadyrov, A.; Abdullayev, S.; Kuttybayeva, A.; Marxuly, S.; Izbairova, A.; Altayeva, Z.; Ermanova, D. Wastewater Treatment Technologies Using Electrical Discharge after Processing of Mineral Raw Materials. Min. Miner. Depos. 2025, 19, 121–131. [Google Scholar] [CrossRef]
  20. Trivedi, Y.; Sharma, M.; Mishra, R.K.; Sharma, A.; Joshi, J.; Gupta, A.B.; Achintya, B.; Shah, K.; Vuppaladadiyamd, A.K. Biochar Potential for Pollutant Removal during Wastewater Treatment: A Comprehensive Review of Separation Mechanisms, Technological Integration, and Process Analysis. Desalination 2025, 600, 118509. [Google Scholar] [CrossRef]
  21. Kamińska, G.; Dave, B.; Appiah, A.N.S.; Majewska, J. Exploitation of Bentonite and Halloysite Fixed Bed Columns—Removal of Organic Micropollutants and Microbiological Regeneration. Desalin. Water Treat. 2025, 322, 101113. [Google Scholar] [CrossRef]
  22. Dave, B.; Moysa, E.L.; Kuźnik, A. Comparative Analysis of Cosmetic Ingredient Degradation: Fungal vs. Bacterial Activity in Diverse Media as Potential Replacements. Int. Biodeterior. Biodegrad. 2024, 191, 105795. [Google Scholar] [CrossRef]
  23. Wei, T.; Zhao, Y.; Zhou, M.; Zhang, Z.; Wei, Y.; Núñez, A.E. Initial Concept and Embodiment to Develop Modular Constructed Wetland: A Unique and Promising Solution to Sustainability Transitions in Water Management. J. Clean. Prod. 2024, 450, 141912. [Google Scholar] [CrossRef]
  24. Apolayo, S.; Deago, E. A Comprehensive Literature Review on Constructed Wetlands: Design, Function, and Modeling Approaches. In Proceedings of the 2024 9th International Engineering, Sciences and Technology Conference (IESTEC), Panama City, Panama, 23–25 October 2024; IEEE: Piscataway, NJ, USA, 2024; pp. 130–133. [Google Scholar]
  25. Wei, J.; Keenahan, J.; Cotterill, S. Review of Experimental Methods and Numerical Models for Hydraulic Studies in Constructed Wetlands. Sustainability 2025, 17, 3303. [Google Scholar] [CrossRef]
  26. Zahra, Z.; Song, M.; Habib, Z.; Ikram, S. Advances in Per- and Polyfluoroalkyl Substances (PFAS) Detection and Removal Techniques from Drinking Water, Their Limitations, and Future Outlooks. Emerg. Contam. 2025, 11, 100434. [Google Scholar] [CrossRef]
  27. Echeverri González, A.F.; Zúñiga-Benítez, H.; Peñuela, G.A. Removal of Herbicide 2,4-D Using Constructed Wetlands at Pilot Scale. Emerg. Contam. 2019, 5, 303–307. [Google Scholar] [CrossRef]
  28. Xu, C.; Feng, Y.; Li, H.; Li, Y.; Yao, Y.; Wang, J. Constructed Wetlands for Mariculture Wastewater Treatment: From Systematic Review to Improvement Measures and Insights. Desalination 2024, 579, 117505. [Google Scholar] [CrossRef]
  29. Jia, W.; Zhang, J.; Wu, J.; Xie, H.; Zhang, B. Effect of Intermittent Operation on Contaminant Removal and Plant Growth in Vertical Flow Constructed Wetlands: A Microcosm Experiment. Desalination 2010, 262, 202–208. [Google Scholar] [CrossRef]
  30. Shelef, O.; Gross, A.; Rachmilevitch, S. Role of Plants in a Constructed Wetland: Current and New Perspectives. Water 2013, 5, 405–419. [Google Scholar] [CrossRef]
  31. Zhang, J.; Shao, Z.; Li, B.; Bai, G.; Yang, L.; Chi, Y.; Wang, M.; Ren, Y. Root Vertical Spatial Stress: A Method for Enhancing Rhizosphere Effect of Plants in Subsurface Flow Constructed Wetland. Environ. Res. 2023, 231, 116083. [Google Scholar] [CrossRef]
  32. Yalcuk, A.; Pakdil, N.B.; Turan, S.Y. Performance Evaluation on the Treatment of Olive Mill Waste Water in Vertical Subsurface Flow Constructed Wetlands. Desalination 2010, 262, 209–214. [Google Scholar] [CrossRef]
  33. Valipour, A.; Ahn, Y.-H. Constructed Wetlands as Sustainable Ecotechnologies in Decentralization Practices: A Review. Environ. Sci. Pollut. Res. 2016, 23, 180–197. [Google Scholar] [CrossRef] [PubMed]
  34. Kumar, S.; Dutta, V. Constructed Wetland Microcosms as Sustainable Technology for Domestic Wastewater Treatment: An Overview. Environ. Sci. Pollut. Res. 2019, 26, 11662–11673. [Google Scholar] [CrossRef] [PubMed]
  35. Fahim, R.; Cheng, L.; Mishra, S. Structural and Functional Perspectives of Carbon Filter Media in Constructed Wetlands for Pollutants Abatement from Wastewater. Chemosphere 2023, 345, 140514. [Google Scholar] [CrossRef]
  36. Zhuang, L.-L.; Li, M.; Li, Y.; Zhang, L.; Xu, X.; Wu, H.; Liang, S.; Su, C.; Zhang, J. The Performance and Mechanism of Biochar-Enhanced Constructed Wetland for Wastewater Treatment. J. Water Process Eng. 2022, 45, 102522. [Google Scholar] [CrossRef]
  37. Feng, L.; Gao, Z.; Hu, T.; He, S.; Liu, Y.; Jiang, J.; Zhao, Q.; Wei, L. Performance and Mechanisms of Biochar-Based Materials Additive in Constructed Wetlands for Enhancing Wastewater Treatment Efficiency: A Review. Chem. Eng. J. 2023, 471, 144772. [Google Scholar] [CrossRef]
  38. Bose, S.; Kumar, P.S.; Vo, D.-V.N.; Rajamohan, N.; Saravanan, R. Microbial Degradation of Recalcitrant Pesticides: A Review. Environ. Chem. Lett. 2021, 19, 3209–3228. [Google Scholar] [CrossRef]
  39. Sharma, P.; Nadda, A.K.; Kumar, S. Microbial Community Profiling in Bio-Stimulated Municipal Solid Waste for Effective Removal of Organic Pollutants Containing Endocrine Disrupting Chemicals. Microbiol. Res. 2023, 267, 127273. [Google Scholar] [CrossRef]
  40. Zhao, C.; Xie, H.; Xu, J.; Xu, X.; Zhang, J.; Hu, Z.; Liu, C.; Liang, S.; Wang, Q.; Wang, J. Bacterial Community Variation and Microbial Mechanism of Triclosan (TCS) Removal by Constructed Wetlands with Different Types of Plants. Sci. Total Environ. 2015, 505, 633–639. [Google Scholar] [CrossRef]
  41. Gupta, P.; Ann, T.; Lee, S.-M. Use of Biochar to Enhance Constructed Wetland Performance in Wastewater Reclamation. Environ. Eng. Res. 2016, 21, 36–44. [Google Scholar] [CrossRef]
  42. Vymazal, J.; Kröpfelová, L. A Three-Stage Experimental Constructed Wetland for Treatment of Domestic Sewage: First 2 Years of Operation. Ecol. Eng. 2011, 37, 90–98. [Google Scholar] [CrossRef]
  43. Stamm, G.; Basapuram, G.; Duttagupta, S.; Dutta, A. Anion-Mediated Pathways in Organophosphate Degradation in the Oconee River Watershed in Georgia. Emerg. Contam. 2025, 11, 100542. [Google Scholar] [CrossRef]
  44. Wang, H.; Sheng, L.; Xu, J. Clogging Mechanisms of Constructed Wetlands: A Critical Review. J. Clean. Prod. 2021, 295, 126455. [Google Scholar] [CrossRef]
  45. Shi, Q.; Yuan, Y.; Zhou, Y.; Yuan, Y.; Liu, L.; Liu, X.; Li, F.; Leng, C.; Wang, H. Pharmaceutical and Personal Care Products (PPCPs) Degradation and Microbial Characteristics of Low-Temperature Operation Combined with Constructed Wetlands. Chemosphere 2023, 341, 140039. [Google Scholar] [CrossRef]
  46. Wu, D.; Van Den Brink, P.J.; Smidt, H.; Bai, H.; Ying, G.-G.; Schuijt, L.M. The Effects of Triclosan on Microbial Communities in Different Sources of Water and Sediments. Ecotoxicol. Environ. Saf. 2025, 303, 118778. [Google Scholar] [CrossRef]
  47. Matamoros, V.; Arias, C.; Brix, H.; Bayona, J.M. Removal of Pharmaceuticals and Personal Care Products (PPCPs) from Urban Wastewater in a Pilot Vertical Flow Constructed Wetland and a Sand Filter. Environ. Sci. Technol. 2007, 41, 8171–8177. [Google Scholar] [CrossRef]
  48. Oh, S.; Choi, D.; Cha, C.-J. Ecological Processes Underpinning Microbial Community Structure during Exposure to Subinhibitory Level of Triclosan. Sci. Rep. 2019, 9, 4598. [Google Scholar] [CrossRef]
  49. Liu, S.; Zhang, Y.; Feng, X.; Pyo, S.-H. Current Problems and Countermeasures of Constructed Wetland for Wastewater Treatment: A Review. J. Water Process Eng. 2024, 57, 104569. [Google Scholar] [CrossRef]
  50. Badhe, N.; Saha, S.; Biswas, R.; Nandy, T. Role of Algal Biofilm in Improving the Performance of Free Surface, up-Flow Constructed Wetland. Bioresour. Technol. 2014, 169, 596–604. [Google Scholar] [CrossRef]
  51. Rosendo, J.C.M.; Da Paz, G.M.; Rosendo, A. Constructed Wetlands Applied on Domestic Wastewater for Decetralized Systems: Concepts, Processes, Modalities, Combinations and Enhancements; a Review. Rev. Environ. Sci. Biotechnol. 2022, 21, 371–397. [Google Scholar] [CrossRef]
  52. Huang, Y.; An, W.; Ning, T.; Ma, Z.; Li, Y.; Liu, K.; Ji, L.; Liu, H.; Hui, D.; Ren, H. Functionality of Bacterial Communities in Constructed Wetlands Used for Water Purification: Influence of Root Components and Seasonality. Front. Plant Sci. 2025, 16, 1480099. [Google Scholar] [CrossRef] [PubMed]
  53. Rodarte-Morales, A.I.; Feijoo, G.; Moreira, M.T.; Lema, J.M. Degradation of Selected Pharmaceutical and Personal Care Products (PPCPs) by White-Rot Fungi. World J. Microbiol. Biotechnol. 2011, 27, 1839–1846. [Google Scholar] [CrossRef]
  54. Alidoosti, F.; Giyahchi, M.; Moghimi, H. Synergistic Bioremediation: Fungal-Bacterial Partnership Degrades LDPE Microplastics Twice as Fast. Curr. Res. Microb. Sci. 2025, 9, 100450. [Google Scholar] [CrossRef] [PubMed]
  55. Łobos-Moysa, E.; Bodzek, M. Application of Hybrid Biological Techniques to the Treatment of Municipal Wastewater Containing Oils and Fats. Desalination Water Treat. 2012, 46, 32–37. [Google Scholar] [CrossRef]
  56. Tu, Y.; Han, L.; Ding, X.; Zhu, X.; Xu, G.; Xie, X.; Zhou, Q.; Shuang, C.; Li, A. Degradation of Pharmaceuticals and Personal Care Products (PPCPs) in Municipal Wastewater by White Rot Fungi: Neutral Environment Exploitation and Degradation Mechanisms. Chem. Eng. J. 2025, 507, 160794. [Google Scholar] [CrossRef]
  57. Khatua, S. Myco-Enzymes as a Potent Tool for the Degradation of Xenobiotics. In Plant-Microbe Interaction Under Xenobiotic Exposure; Roy, S., Mandal, V., Eds.; Springer Nature Singapore: Singapore, 2025; pp. 549–584. ISBN 978-981-96-8259-1. [Google Scholar]
  58. Upadhyay, R.; Przystaś, W.; Dave, B. Myco-Remediation of Synthetic Dyes: A Comprehensive Review on Contaminant Alleviation Mechanism, Kinetic Study and Toxicity Analysis. Int. J. Environ. Sci. Technol. 2025, 22, 521–538. [Google Scholar] [CrossRef]
  59. Nie, H.; Guo, D.; Wang, Y.; Wang, L.; Cao, S.; Huang, Z.; Zhao, T. Glucose Co-Metabolism Enhances Extracellular ROS-Mediated TBBPA Biodegradation by Pseudomonas Aeruginosa NY3 and Reshapes Microbial Communities in Wastewater Treatment Systems. J. Water Process Eng. 2025, 75, 107989. [Google Scholar] [CrossRef]
  60. Das, P.; Mehra, A.; Sachan, S.G.; Chattopadhyay, S. Screening Different Solid Supports for Pseudomonas Aeruginosa Biofilm Formation and Determining Its Efficiency for Decolorization and Degradation of Congo Red. Arch. Microbiol. 2024, 206, 402. [Google Scholar] [CrossRef]
  61. Jureczko, M.; Przystaś, W.; Krawczyk, T.; Gonciarz, W.; Rudnicka, K. White-Rot Fungi-Mediated Biodegradation of Cytostatic Drugs—Bleomycin and Vincristine. J. Hazard. Mater. 2021, 407, 124632. [Google Scholar] [CrossRef]
  62. Javid, F.; Ali, G.; Rehman, A.; Naeem, R.; Ali, I.; Naz, I. Assessment of Plastic Degradation by Indigenous Bacteria from Waste Disposal Sites. Emerg. Contam. 2024, 10, 100323. [Google Scholar] [CrossRef]
  63. Dave, B.; Moysa, E.L.; Kuźnik, A. Nicotine Degradation by Trametes Versicolor: Insights from Diverse Environmental Stressors and Wastewater Medium. Molecules 2025, 30, 2658. [Google Scholar] [CrossRef]
  64. Younes, E.A.; El-Sheikh, A.H.; Alsmadi, R.B. The Use of New Class α-Amino Nitrile Modified Magnetic Adsorbents for Removal of Cd(II) from Aqueous Medium: Sorbent Modification vs. α-Amino Nitrile Addition to the Adsorption Medium. Emerg. Contam. 2024, 10, 100261. [Google Scholar] [CrossRef]
  65. Rodrigues, R.; Oliveira, M.B.P.P.; Alves, R.C. Chlorogenic Acids and Caffeine from Coffee By-Products: A Review on Skincare Applications. Cosmetics 2023, 10, 12. [Google Scholar] [CrossRef]
  66. Gautam, P.; Carsella, J.S.; Kinney, C.A. Presence and Transport of the Antimicrobials Triclocarban and Triclosan in a Wastewater-Dominated Stream and Freshwater Environment. Water Res. 2014, 48, 247–256. [Google Scholar] [CrossRef] [PubMed]
  67. Schabacker, J.; Gerlach, J.; Münderle, M.; Dietzen, C.; Ludwigs, J.-D. Focal Bird Species and Risk Assessment Approach for Nonagricultural Grassland Scenarios in Central Europe. Environ. Toxicol. Chem. 2014, 33, 2055–2061. [Google Scholar] [CrossRef] [PubMed]
  68. Adhikari, S.; Kumar, R.; Driver, E.M.; Perleberg, T.D.; Yanez, A.; Johnston, B.; Halden, R.U. Mass Trends of Parabens, Triclocarban and Triclosan in Arizona Wastewater Collected after the 2017 FDA Ban on Antimicrobials and during the COVID-19 Pandemic. Water Res. 2022, 222, 118894. [Google Scholar] [CrossRef]
  69. Marco-Urrea, E.; Pérez-Trujillo, M.; Cruz-Morató, C.; Caminal, G.; Vicent, T. Degradation of the Drug Sodium Diclofenac by Trametes Versicolor Pellets and Identification of Some Intermediates by NMR. J. Hazard. Mater. 2010, 176, 836–842. [Google Scholar] [CrossRef]
  70. Bhattacherjee, A.K.; Shukla, P.K.; Dikshit, A.; Garg, N. Biodegradation of Imidacloprid by Microbial Consortium in Sterile and Non-Sterile Soil under in Vitro Conditions. J. Eco-Friendly Agric. 2022, 17, 105–109. [Google Scholar] [CrossRef]
  71. Wdowczyk, A.; Szymańska-Pulikowska, A. Effect of Substrates on the Potential of Phragmites Australis to Accumulate and Translocate Selected Contaminants from Landfill Leachate. Water Resour. Ind. 2023, 29, 100203. [Google Scholar] [CrossRef]
  72. Truu, M.; Juhanson, J.; Truu, J. Microbial Biomass, Activity and Community Composition in Constructed Wetlands. Sci. Total Environ. 2009, 407, 3958–3971. [Google Scholar] [CrossRef]
  73. Dave, B.; Sanghvi, G. Enzyme Action for Dye Degradation. In Dye Biodegradation, Mechanisms and Techniques; Muthu, S.S., Khadir, A., Eds.; Sustainable Textiles: Production, Processing, Manufacturing & Chemistry; Springer Singapore: Singapore, 2022; pp. 141–163. ISBN 978-981-16-5931-7. [Google Scholar]
  74. Keizer-Vlek, H.E.; Verdonschot, P.F.M.; Verdonschot, R.C.M.; Dekkers, D. The Contribution of Plant Uptake to Nutrient Removal by Floating Treatment Wetlands. Ecol. Eng. 2014, 73, 684–690. [Google Scholar] [CrossRef]
  75. Vymazal, J. Constructed Wetlands for Wastewater Treatment: Five Decades of Experience. Environ. Sci. Technol. 2011, 45, 61–69. [Google Scholar] [CrossRef] [PubMed]
  76. Kadlec, R.H.; Wallace, S. Treatment Wetlands, 2nd ed.; CRC Press: Boca Raton, FL, USA, 2008; ISBN 978-0-429-13795-2. [Google Scholar]
  77. Ilyas, H.; Van Hullebusch, E.D. Performance Comparison of Different Constructed Wetlands Designs for the Removal of Personal Care Products. Int. J. Environ. Res. Public Health 2020, 17, 3091. [Google Scholar] [CrossRef] [PubMed]
  78. Löffler, P.; Escher, B.I.; Baduel, C.; Virta, M.P.; Lai, F.Y. Antimicrobial Transformation Products in the Aquatic Environment: Global Occurrence, Ecotoxicological Risks, and Potential of Antibiotic Resistance. Environ. Sci. Technol. 2023, 57, 9474–9494. [Google Scholar] [CrossRef]
  79. Faulwetter, J.L.; Gagnon, V.; Sundberg, C.; Chazarenc, F.; Burr, M.D.; Brisson, J.; Camper, A.K.; Stein, O.R. Microbial Processes Influencing Performance of Treatment Wetlands: A Review. Ecol. Eng. 2009, 35, 987–1004. [Google Scholar] [CrossRef]
  80. Gruenke, L.D.; Craig, J.C.; Wester, R.C.; Maibach, H.I.; North-Root, H.; Corbin, N.C. A Selected Ion Monitoring GC/MS Assay for 3,4,4′-Trichlorocarbanilide and Its Metabolites in Biological Fluids. J. Anal. Toxicol. 1987, 11, 75–80. [Google Scholar] [CrossRef]
  81. Kennedy, R.; Fling, R.; Terry, P.; Menn, F.-M.; Chen, J.; Borman, C. Extraction of 3,4,4′-Trichlorocarbanilide from Rat Fecal Samples for Determination by High Pressure Liquid Chromatography–Tandem Mass Spectrometry. Int. J. Environ. Res. Public Health 2015, 12, 8125–8132. [Google Scholar] [CrossRef]
  82. Papadimitriou, C.A.; Papatheodoulou, A.; Takavakoglou, V.; Zdragas, A.; Samaras, P.; Sakellaropoulos, G.P.; Lazaridou, M.; Zalidis, G. Investigation of Protozoa as Indicators of Wastewater Treatment Efficiency in Constructed Wetlands. Desalination 2010, 250, 378–382. [Google Scholar] [CrossRef]
  83. Akhiladas, E.K.; Chakraborty, S. Bioremediation of Petroleum Refinery Wastewater Using Constructed Wetlands: A Focus on Microbial Diversity and Pollutant Degradation. Int. Biodeterior. Biodegrad. 2025, 204, 106152. [Google Scholar] [CrossRef]
  84. Rani, A.; Chauhan, M.; Kumar Sharma, P.; Kumari, M.; Mitra, D.; Joshi, S. Microbiological Dimensions and Functions in Constructed Wetlands: A Review. Curr. Res. Microb. Sci. 2024, 7, 100311. [Google Scholar] [CrossRef]
  85. Ali, H.; Min, Y.; Yu, X.; Kooch, Y.; Marnn, P.; Ahmed, S. Composition of the Microbial Community in Surface Flow-Constructed Wetlands for Wastewater Treatment. Front. Microbiol. 2024, 15, 1421094. [Google Scholar] [CrossRef]
  86. Nivala, J.; Wallace, S.; Van Afferden, M.; Müller, R.A. Evapotranspiration Dynamics in Aerated and Non-Aerated Subsurface Flow Treatment Wetlands. Sci. Total Environ. 2022, 843, 156605. [Google Scholar] [CrossRef]
  87. Wang, H.; Xu, D.; Han, J.; Xu, R.; Han, D. Reshaped Structure of Microbial Community within a Subsurface Flow Constructed Wetland Response to the Increased Water Temperature: Improving Low-Temperature Performance by Coupling of Water-Source Heat Pump. Sci. Total Environ. 2021, 781, 146798. [Google Scholar] [CrossRef]
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Figure 1. Wetland setup formation and aeration mechanism.
Figure 1. Wetland setup formation and aeration mechanism.
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Figure 2. (a) Mycelium growth inhibition (incubation 25°), (b) inhibition of bacterial growth (incubation 37°).
Figure 2. (a) Mycelium growth inhibition (incubation 25°), (b) inhibition of bacterial growth (incubation 37°).
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Figure 3. (a) Fungal and bacterial biomass growth, (b) plant growth.
Figure 3. (a) Fungal and bacterial biomass growth, (b) plant growth.
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Figure 4. pH levels during experiment time in the wetland setup.
Figure 4. pH levels during experiment time in the wetland setup.
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Figure 5. SEM images of stones, (a) before experiment, (b–d) bacterial accumulation after experiment, (e) edge of the stone.
Figure 5. SEM images of stones, (a) before experiment, (b–d) bacterial accumulation after experiment, (e) edge of the stone.
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Figure 6. (a) 1H NMR spectrum of wetland control, (b) 13C NMR spectrum of wetland control, 1H NMR spectrum of 4 weeks (c) and 7 weeks (d) of experiment from wetland (bacteria + fungi).
Figure 6. (a) 1H NMR spectrum of wetland control, (b) 13C NMR spectrum of wetland control, 1H NMR spectrum of 4 weeks (c) and 7 weeks (d) of experiment from wetland (bacteria + fungi).
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Figure 7. 1H NMR spectrum of 4 weeks (a) and 7 weeks (b) of experiment from wetland B (fungi), 1H NMR spectrum of 4 weeks (c) and 7 weeks (d) of experiment form wetland A (bacteria).
Figure 7. 1H NMR spectrum of 4 weeks (a) and 7 weeks (b) of experiment from wetland B (fungi), 1H NMR spectrum of 4 weeks (c) and 7 weeks (d) of experiment form wetland A (bacteria).
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Table 1. Summary and composition of wetland setup.
Table 1. Summary and composition of wetland setup.
ParametersWetland A (Bacteria)Wetland B (Fungi)Wetland C (Bacteria and Fungi)Wetland D (Control)
Capacity30 L30 L30 L30 L
Length (cm)40 cm40 cm40 cm40 cm
Width (cm)25 cm25 cm25 cm25 cm
Height (cm)30 cm30 cm30 cm30 cm
Synthetic wastewater volume15 L15 L15 L15 L
Porosity (%)40–4540–4350–5250–55
Heater and thermostatyesyesyesyes
Temperature37 °C25 °C30 °CRoom temp.
Microbial speciesPseudomonas aeruginosaTrametes versicolorPseudomonas aeruginosa
+
Trametes versicolor		None
Aeration setupyesnoyesyes
Vegetation typeSpecies Phragmites australisSpecies Phragmites australisSpecies Phragmites australisSpecies Phragmites australis
Table 2. NMR baseline setup of pollutants and chemical structure of all pollutants.
Table 2. NMR baseline setup of pollutants and chemical structure of all pollutants.
PollutantBaseline 1H NMR Characteristic Peak (δ in ppm)Initial Degradation (% at t = 0)
Caffeine(7.94) (A prominent singlet)(0%) (Figure 6a,b)
Methylparaben(2.19) (Methyl proton peak)(0%) (Figure 6a,b)
Trichlorocarbanilide (TCC)Not detectableNot applicable
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Table 3. Degradation result of the wetland experiment.
Table 3. Degradation result of the wetland experiment.
Treatment (Wetland)Time (Weeks)Caffeine Degradation %Methylparaben
Degradation %		TCC Degradation %Key Observation
Wetland D controlFirst dayPeak at 7.94, 0% degradation
(Figure 6a,b)		Peak at 2.19, 0% degradation
(Figure 6a,b)		Not detectable
(Figure 6a,b)		-
Wetland A (fungi)4th weekVery small peak, 95 to 99% degradation
(Figure 7a)		Peak value at 1.62, 73.97% degradation
(Figure 7a)		-High initial caffeine removal
7th weekNo value, 99 to 100% degradation
(Figure 7b)		No value, 99 to 100% degradation
(Figure 7b)		-Complete removal achieved
Wetland B (bacteria)4th weekPeak value at 2.36, 70.28% degradation
(Figure 7c)		Peak value at 1.58, 72.14% degradation
(Figure 7c)		-Slower initial removal rate
7th weekNo value, 99 to 100% degradation
(Figure 7d)		No value, 99 to 100% degradation
(Figure 7d)		-Complete removal achieved
Wetland C
(fungi + bacteria)		4th weekNo value, 99 to 100% degradation
(Figure 6c)		No value, 99 to 100% degradation
(Figure 6c)		-Fastest Removal and total degradation
7th weekNo value, 99 to 100% degradation
(Figure 6d)		No value, 99 to 100% degradation
(Figure 6d)		-
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Dave, B.; Łobos-Moysa, E.; Kuznik, A.; Maqsood, A.; Appiah, A.N.S.; Krzeszowski, S.; Joshi, R. Enhancing Microbial Biodegradation of PPCPs in Wastewater via Natural Self-Purification in a Novel Constructed Wetland System. Sustainability 2026, 18, 548. https://doi.org/10.3390/su18010548

AMA Style

Dave B, Łobos-Moysa E, Kuznik A, Maqsood A, Appiah ANS, Krzeszowski S, Joshi R. Enhancing Microbial Biodegradation of PPCPs in Wastewater via Natural Self-Purification in a Novel Constructed Wetland System. Sustainability. 2026; 18(1):548. https://doi.org/10.3390/su18010548

Chicago/Turabian Style

Dave, Bhautik, Ewa Łobos-Moysa, Anna Kuznik, Abdullah Maqsood, Augustine Nana Sekyi Appiah, Swiatoslaw Krzeszowski, and Rushikesh Joshi. 2026. "Enhancing Microbial Biodegradation of PPCPs in Wastewater via Natural Self-Purification in a Novel Constructed Wetland System" Sustainability 18, no. 1: 548. https://doi.org/10.3390/su18010548

APA Style

Dave, B., Łobos-Moysa, E., Kuznik, A., Maqsood, A., Appiah, A. N. S., Krzeszowski, S., & Joshi, R. (2026). Enhancing Microbial Biodegradation of PPCPs in Wastewater via Natural Self-Purification in a Novel Constructed Wetland System. Sustainability, 18(1), 548. https://doi.org/10.3390/su18010548

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