Nature-based bioreactors: Tackling antibiotic resistance in urban wastewater treatment.

The overuse and misuse of antibiotics have accelerated the selection of antibiotic-resistant bacteria, significantly impacting human, animal, and environmental health. As aquatic environments are vulnerable to antibiotic resistance, suitable management practices should be adopted to tackle this phenomenon. Here we show an effective, nature-based solution for reducing antibiotic resistance from actual wastewater. We utilize a bioreactor that relies on benthic (biofilms) and planktonic microbial communities to treat secondary effluent from a small urban wastewater treatment plant (<10,000 population equivalent). This treated effluent is eventually released into the local aquatic ecosystem. We observe high removal efficiency for genes that provide resistance to commonly used antibiotic families, as well as for mobile genetic elements that could potentially aid in their spread. Importantly, we notice a buildup of sulfonamide (sul1 and sul2) and tetracycline (tet(C), tet(G), and tetR) resistance genes specifically in biofilms. This advancement marks the initial step in considering this bioreactor as a nature-based, cost-effective tertiary treatment option for small UWWTPs facing antibiotic resistance challenges.

Exposure and recovery: The effect of different dilution factors of treated and untreated metal mining effluent on freshwater biofilm function and structure.

Abandoned mines generate effluents rich in heavy metals, and these contaminants are released uncontrolled into the nearby aquatic ecosystems, causing severe pollution. However, no real solution exists, leaving a legacy of global pollution. In this study, the efficiency of the treatment technologies in reducing the ecological impacts of mining effluents to freshwater ecosystems with different dilution capacities was tested using biofilm communities as biological indicators. The functional and structural recovery capacity of biofilm communities after 21 days of exposure was assessed. With this aim, we sampled aquatic biofilms from a pristine stream and exposed them to treated (T) and untreated (U) metal mining effluent from Frongoch abandoned mine (Mid Wales, UK). Additionally, we simulated two different flow conditions for the receiving stream: high dilution (HD) and low dilution (LD). After exposure, the artificial streams were filled with artificial water for 14 days to assess the biofilm recovery. Unexposed biofilm served as control for biofilm responses (functional and structural) measured throughout time. During the exposure, short term effects on biofilm functioning (photosynthetic efficiency, nutrient uptake) were observed in T-LD, U-HD, and U-LD, whereas long term effects (community composition, chl-a, and diatom metrics) were observed on the structure of all biofilms exposed to the treated and untreated mining effluent. On the other hand, metal accumulation occurred in biofilms exposed to the mining effluents. However, a functional recovery was observed for all treatments, except in the U-LD in which biofilm structure did not present a significant recovery after the exposure period. The results presented here highlight the need to consider the dilution capacity of the receiving stream to assess the real efficiency of treatment technologies applied to mining effluents to mitigate the ecological impact on freshwater ecosystems.

What happens when salinization meets eutrophication? A test using stream microcosms.

Nutrient and salt pollution often co-occur in rivers and streams due to human activities (e.g., agriculture, urbanization). Thus, understanding the interactive effects of nutrients and salinity on freshwater ecosystems is critical for environmental management. We experimentally assessed the interactive effects of nutrient and salt pollution on stream microcosms using biofilm and macroinvertebrates as model systems. Six treatments were performed in triplicate: control (C: N-NH4+ = 0.05; P- PO43- = 0.037; Cl- = 33.5 mg L-1), intermediate nutrient (IN: N-NH4+ = 0.4; P- PO43- = 0.271; Cl- = 33. 5 mg L-1), high nutrient (HN: N-NH4+ = 0.84; P- PO43- = 0.80; Cl- = 33.5 mg L-1), salt (S: N-NH4+ = 0.05; P- PO43- = 0.037; Cl- = 3000 mg L-1), salt with intermediate nutrient (SIN: N-NH4+ = 0.4; P- PO43- = 0.27; Cl- = 3000 mg L-1) and salt with high nutrient (SHN: N-NH4+ = 0.84; P- PO43- = 0.80; Cl- = 3000 mg L-1). After 14 days of exposure, biofilm chlorophyll-a increased across all treatments, with cyanobacteria replacing diatoms and green algae. Treatments with no added nutrients (C and S) had more P uptake capacity than the rest. The indicator species analysis showed 8 significant taxa, with Orthocladius (Orthocladius) gr. Wetterensis and Virganytarsus significantly associated with the salinity treatment. Overall, salt pollution led to a very strong decline in macroinvertebrate richness and diversity. However, salt toxicity seemed to be ameliorated by nutrient addition. Finally, both structural equation models and biotic-abiotic interaction networks showed that complex biological interactions could be modulating the response of the biological communities to our treatments. Thus, our study calls for species-level assessments of salt and nutrient effects on river ecosystems and advocates for better management of co-occurring pollutants.

Dissipation of pesticides by stream biofilms is influenced by hydrological histories.

To evaluate the effects of hydrological variability on pesticide dissipation capacity by stream biofilms, we conducted a microcosm study. We exposed biofilms to short and frequent droughts (daily frequency), long and less frequent droughts (weekly frequency) and permanently immersed controls, prior to test their capacities to dissipate a cocktail of pesticides composed of tebuconazole, terbuthylazine, imidacloprid, glyphosate and its metabolite aminomethylphosphonic acid. A range of structural and functional descriptors of biofilms (algal and bacterial biomass, extracellular polymeric matrix (EPS) concentration, microbial respiration, phosphorus uptake and community-level physiological profiles) were measured to assess drought effects. In addition, various parameters were measured to characterise the dynamics of pesticide dissipation by biofilms in the different hydrological treatments (% dissipation, peak asymmetry, bioconcentration factor, among others). Results showed higher pesticide dissipation rates in biofilms exposed to short and frequent droughts, despite of their lower biomass and EPS concentration, compared to biofilms in immersed controls or exposed to long and less frequent droughts. High accumulation of hydrophobic pesticides (tebuconazole and terbuthylazine) was measured in biofilms despite the short exposure time (few minutes) in our open-flow microcosm approach. This research demonstrated the stream biofilms capacity to adsorb hydrophobic pesticides even in stressed drought environments.