Size-dependent effects of microplastics on intestinal microbiome for Perna viridis.

Microplastics pollution threatens to marine organisms, particularly bivalves that actively ingest and accumulate microplastics of certain sizes, potentially disrupting intestinal homeostasis. This study investigated the microplastic abundance in wild and farmed mussels around Singapore, and examined the size-dependent effects of nano- to micro-scale polystyrene (0.5 µm/5 µm/50 µm) on the mussel intestinal microbiome in the laboratory. The field investigation revealed higher microplastic abundance in farmed mussels compared to wild ones. Experimentally, mussels exposed to 0.6 mg/L of microplastics for 7 days, followed by a 7-day depuration period, showed substantial impacts on Spirochaetes and Proteobacteria, facilitating the proliferation of pathogenic species and differentially affecting their pathogenic contributions. Metagenomics analysis revealed that microplastic exposure reduced Spirochaeta's contribution to virulence and pathogenicity loss, did not affect Vibrio and Oceanispirochaeta's pathogenicity, and increased Treponema and Oceanispirochaeta's contributions to pathogenicity loss. Moreover, microplastics increased transmembrane transporters and impacted oxidative phosphorylation enzymes, impairing energy metabolism. These effects persisted after depuration, indicating lack of resilience in the microbiome. Nano- and micro-scale plastics perturbed the mussel microbiome composition and functions in a size-dependent manner, with nano-plastics being the most disruptive. The increasing use and sale of aquaculture equipment of plastic may exacerbate the intestinal dysbiosis in bivalves, which threatens consumers' health.

Integrated metagenomic and metaproteomic analyses reveal bacterial micro-ecological mechanisms in coral bleaching.

IMPORTANCE: Coral reefs worldwide are facing rapid decline due to coral bleaching. However, knowledge of the physiological characteristics and molecular mechanisms of coral symbionts respond to stress is scarce. Here, metagenomic and metaproteomic approaches were utilized to shed light on the changes in the composition and functions of coral symbiotic bacteria during coral bleaching. The results demonstrated that coral bleaching significantly affected the composition of symbionts, with bacterial communities dominating in bleached corals. Through differential analyses of gene and protein expression, it becomes evident that symbionts experience functional disturbances in response to heat stress. These disturbances result in abnormal energy metabolism, which could potentially compromise the health and resilience of the symbionts. Furthermore, our findings highlighted the highly diverse microbial communities of coral symbionts, with beneficial bacteria providing critical services to corals in stress responses and pathogenic bacteria driving coral bleaching. This study provides comprehensive insights into the complex response mechanisms of coral symbionts under heat stress from the micro-ecological perspective and offers fundamental data for future monitoring of coral health.

Size-dominated biotoxicity of microplastics laden with benzophenone-3 and ciprofloxacin: Enhanced integrated biomarker evaluation on mussels.

Microplastics (MPs) are emerging pollutants with diverse sizes in aquatic environments. This paper investigates the toxicity of micron- and nano-scale polystyrene (50 µm, 5 µm, 0.5 µm) loaded with 2-hydroxy-4-methoxy-benzophenone (BP-3) and ciprofloxacin (CIP) by eight biomarker responses in mussels, perna viridis. The mussels were exposed to MPs and chemicals for 7 days before 7 days of depuration. Eight biomarkers were measured to determine biotoxicity over time by using the weighted integrated biomarkers index evaluation (EIBR). Mussels exposed to MPs on a daily basis demonstrated a cumulative toxic effect. The toxicity of MPs for mussels was inversely related to the size at which they can be ingested. Then toxicity was reversed when exposure was halted. EIBR mold has shown a significant difference in the biotoxicity of each biological level under different exposure scenarios. In general, the mussel toxicity influenced by BP-3 and CIP exposure without an adsorbent was insignificant. MPs laden with them increased the toxicity of mussels. Under condition of lower concentration of ECs (Emerging contaminants), the presence of MPs as a component of a combined pollutant in water dominated the biotoxicity for mussels. The EIBR assessment further validated that the biotoxicity of mussels was size-dependent. Its application simplified the biomarkers' response index and enhanced the accuracy of evaluation by weighing on molecular, cellular and physiological level. Specifically, mussels were physiologically sensitive to nano-scale plastics, with nano-scale plastics causing a higher level of cellular immunity destruction and genotoxicity than micron-scale plastics. Enzymatic antioxidant systemswere upregulated based on size-differential plastics; however, the total antioxidant effect of non-enzymatic defenses appeared to be least affected by the size effect.

Holes on surfaces of the weathered plastic fragments from coastal beaches.

The surface morphology of weathered plastics undergoes a variety of changes. In this study, 3950 plastic fragments from 26 beaches around the world, were assessed to identify holes. Holes were identified on 123 fragments on 20 beaches, with the highest frequency (10.3 %) being identified at Qesm AL Gomrok Beach in Egypt. The distribution of holes could be divided into even, single-sided, and random types. The external and internal holes were similar in size (37 ± 15 µm) of even type fragments. The external holes were larger than the internal holes in single-sided (516 ± 259 µm and 383 ± 161 µm) and random (588 ± 262 µm and 454 ± 210 µm) fragment types. The external hole sizes were positively correlated with the internal hole sizes for each type. This study reports a novel deformation phenomenon on the surface of weathered plastics and highlights their potential effects on plastics.

Bacterial Communities Vary from Different Scleractinian Coral Species and between Bleached and Non-Bleached Corals.

Bleaching is one of the most relevant factors implicated in the integrity of coral reef ecosystems, with the increasing frequency and intensity of damaging events representing a serious threat to reef biodiversity. Here, we analyzed changes in coral-associated bacteria from three types of non-bleached and bleached scleractinian corals (Acropora digitifera, Galaxea fascicularis, and Porites pukoensis) in Hainan Luhuitou peninsula coastal areas. The community structure of symbiotic bacteria differed significantly among the three apparently healthy corals. The bleached corals had higher bacterial alpha diversity and some specific bacteria genera, including Ruegeria, Methyloceanibacter, Filomicrobium, Halioglobus, Rubripirellula, Rhodopirellula, Silicimonas, Blastopirellula, Sva0996 marine group, Woeseia, and unclassified_c_Gammaproteobacteria, were consistently increased in bleached groups. Network analysis revealed significantly different degrees of modularity between bleached and non-bleached groups at the bacterial genus level, and a higher proportion of links was dominated by positive co-occurrences. Functional prediction analysis illustrated that coral-associated bacteria remained relatively consistent in the bleached and non-bleached groups. Structure equation modeling revealed that the bacterial community diversity and function were directly influenced by host and environment factors. These findings suggested that coral-associated bacterial responses to bleaching occur in a host-dependent manner, informing novel strategies for restoring coral and aiding adaption to bleaching stress. IMPORTANCE Accumulating evidence indicates that coral-associated bacteria play an important role in the health of holobionts. However, the variability of the symbiotic bacterial community structure among coral species with different coral health statuses remains largely unknown. Here, we investigated three apparent non-bleached (healthy) and bleached coral species (sampled in situ), involving related symbiotic bacterial profiles, including composition, alpha diversity, network relationship, and potential function. Structural equation modeling analysis was used to analyze the relationship between coral status and abiotic and biotic factors. The bacterial community structure of different groups was shown to exhibit host-specific traits. Both host and environmental impacts had primary effects on coral-associated microbial communities. Future studies are needed to identify the mechanisms that mediate divergent microbial consortia.

Process-oriented impacts of microplastic fibers on behavior and histology of fish.

Microplastic pollution has raised global concern for its hazards to biota. To determine the direct impact of microplastics during their contact with fish, we exposed goldfish (Carassius auratus) to 100 and 1000 items/L waterborne microplastic fibers in the short- and long-term. In the presence of 1000 items/L of microplastic fibers, the coughing behavior of fish increased significantly after 2 h of exposure. Predatory behaviors decreased significantly by 53.0% after 45 d of exposure, and the reduction in daily food intake was negatively related to exposure duration in the 1000 items/L group. In addition, microplastic fibers stimulated dynamic mucus secretion across different fish tissues during the different processes evaluated in this study, with 30.0% and 62.9% overall increases in the secretory capacity of mucus cells in the 100 and 1000 items/L groups, respectively. These behavioral and histological alterations were derived from the ventilation, feeding, and swimming processes of goldfish. We regarded these changes as process-oriented impacts, suggesting the effects of microplastics on fish and how fish cope with microplastics.

Wastewater surveillance of SARS-CoV-2 and chemical markers in campus dormitories in an evolving COVID - 19 pandemic.

In this study, we report the implementation of a comprehensive wastewater surveillance testing program at a university campus in Singapore to identify Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) infected individuals and the usage of pharmaceuticals and personal care products (PPCPs) as well as other emerging contaminants (ECs). This unique co-monitoring program simultaneously measured SARS-CoV-2 with chemical markers/contaminants as the COVID-19 situation evolved from pandemic to endemic stages, following a nationwide mass vaccination drive. SARS-CoV-2 RNA concentrations in wastewater from campus dormitories were measured using real-time reverse transcription-polymerase chain reaction (RT-qPCR) and corroborated with the number of symptomatic COVID-19 cases confirmed with the antigen rapid test (ART). Consistent results were observed where the concentrations of SARS-CoV-2 RNA detected in wastewater increased proportionately with the number of COVID-19 infected individuals residing on campus. Similarly, a wide range of ECs, including disinfectants and antibiotics, were detected through sensitive liquid chromatography with tandem mass spectrometry (LC-MS/MS) techniques to establish PPCPs consumption patterns during various stages of the COVID-19 pandemic in Singapore. Statistical correlation of SARS-CoV-2 RNA was observed with few ECs belonging to disinfectants, PCPs and antibiotics. A high concentration of disinfectants and subsequent positive correlation with the number of reported cases on the university campus indicates that disinfectants could serve as a chemical marker during such unprecedented times.

Prokaryotic and eukaryotic microbial communities associated with coral species have high host specificity in the South China Sea.

Reef-building corals are well known for their obligate association with Symbiodiniaceae, and an array of other microbes, including bacteria, fungi, and symbiotic algae (i.e., total microbiome), which together form the coral holobiont. The total microbiome plays an intricate part in maintaining the homeostasis of the coral holobiont and is closely associated with host health. However, the composition of the coral associated microbiome and interaction between its different members remains elusive because few analyses have bridged taxonomically disparate groups. This research gaps have prevented a holistic understanding of the total microbiome. Thus, to simultaneously characterize the bacterial, fungal and symbiotic algal communities associated with different coral species, and explore the relationship between these symbionts and coral health, healthy and bleached tissues from four coral species, Acropora muricata, Galaxea fascicularis, Platygyra daedalea, and Pavona explanulata, were collected from the Xisha Islands of the South China Sea. Using high throughput sequencing, a high degree of host-specificity was observed among bacterial, fungal, and algal groups across coral species. There were no obvious changes in the microbial community structure of apparently healthy and bleached corals, but host bleaching allowed colonization of the holobionts by diverse opportunistic microbes, resulting in a significant elevation in the α-diversity of microbial communities. In addition, co-occurrence analysis of the coral microbiota also identified more complex microbial interactions in bleached corals than in healthy ones. In summary, this study characterized the structure of coral-associated microbiomes across four coral species, and systematically studied microbiome differences between healthy and bleached corals. The findings improve our understanding of the heterogeneity of symbiotic microorganisms and the impact of coral's physiological status on its associated microbial communities composition.

Assessment of human exposure to benzophenone-type UV filters: A review.

To avoid the harmful effects of UV radiation, benzophenone-type UV filters (BPs) are widely used in personal care products and other synthetic products. Biomonitoring studies have shown the presence of BPs in various human biological samples, raising health concerns. However, there is a paucity of data on the global human exposure to this group of contaminants. In this study, we compiled data on the body burden of BPs along with the possible exposure routes and biotransformation pathways. BPs can easily penetrate the skin barrier and thus, they can be absorbed through the skin. In the human body, BPs can undergo Phase I (mainly demethylation and hydroxylation) and Phase II (mainly glucuronidation and sulfation) biotransformations. From a total of 158 studies, most of the studies are related to urine (concentration up to 92.7 mg L-1), followed by those reported in blood (up to 0.9 mg L-1) and milk (up to 0.8 mg L-1). Among BPs, benzophenone-1 and benzophenone-3 are the most commonly detected congeners. The body burden of BPs is associated with various factors, including the country of residence, lifestyle, income, education level, and ethnicity. The presence of BPs in maternal urine (up to 1.1 mg L-1), placenta (up to 9.8 ng g-1), and amniotic fluid (up to 15.7 µg L-1) suggests potential risks of prenatal exposure. In addition, transplacental transfer of BPs is possible, as demonstrated by their presence in maternal serum and cord serum. The possible association of BPs exposure and health effects was discussed. Future human biomonitoring studies and studies on the potential health effects are warranted. Overall, this review provides a summary of the global human exposure to BPs and can serve as supporting evidence to guide usage in order to protect humans from being exposed to BPs.

Methanosarcina thermophila bioaugmentation and its synergy with biochar growth support particles versus polypropylene microplastics in thermophilic food waste anaerobic digestion.

Both biochar supplementation as well as bioaugmentation have been shown in literature to improve the methane yield of anaerobic digestion. In this study, the combination of both are evaluated by growing Methanosarcina thermophila on biochar support particles prior to augmentation of thermophilic food waste anaerobic digestion. Biochar stand alone, bioaugmentation solely, a combination of both added separately or grown together, and utilizing polypropylene (PP) microplastics as growth support instead were all tested when starting up a thermophilic process from mesophilic inoculum. Methanosarcina thermophila and biochar supplementation displayed synergy, with 5% M. thermophila on 1 g/L biochar presenting a 32% increase in specific methane yield over the control. Double the bioaugmentation dosage/concentration was also trialled with a thermophilic inoculum, and 10% M. thermophila grown on 2 g/L biochar displayed the best results with a 20% increase specific methane yield from its control standard.

Impacts of size-fractionation on toxicity of marine microplastics: Enhanced integrated biomarker assessment in the tropical mussels, Perna viridis.

Accumulation of microplastics (MP) in oceanic waters is eroding the health of marine biota. We investigated how size-fractionated MP influence the toxicity risks towards a tropical keystone species, Perna viridis. Tissue-specific bioaccumulation and in vivo toxicity of polystyrene (PS) particles (0.5, 5, and 50 µm) were measured upon continuous exposure for 7 days, followed by 7 days depuration. P. viridis were exposed to equivalent mass (0.6 mg/L), corresponding to 4.0-4.6 particles/mL, 4.6-7.1 × 103 particles/mL, and 1.1-4.8 × 106 particles/mL for 50 µm, 5 µm and 0.5 µm PS particles, respectively. Onset toxicity were quantified through the enhanced integrated multi-biomarker response (EIBR) model, measured by weighting of biological organisation levels of eight biomarkers: (i) molecular (i.e., DNA damage (comet), 7-ethoxy resorufin O-deethylase (EROD), Catalase (CAT), Superoxide dismutase (SOD), Ferric Reducing Antioxidant Power (FRAP)); (ii) cellular (i.e., Neutral red retention (NRR), phagocytosis); and (iii) physiological (i.e., filtration rate). Data showed slightly elevated lysosomal instability (NRR) and antioxidant defences (FRAP, SOD, CAT, EROD) in specimens exposed to nano-PS (0.5 µm) compared to micro-PS (5 and 50 µm). Immunotoxicity (phagocytosis) and genotoxicity (comet) for haemocyte cells were significantly higher in specimens exposed to nano-PS (p < 0.05). EIBR index corroborated increasing toxicity modulated by MP sizes in descending order: 0.5 µm > 5 µm > 50 µm, with nano-PS exerted significantly higher biological effects (EIBR = 19.77 ± 5.89) than the unexposed group (EIBR = 10.97 ± 2.02; p < 0.05). Symptomatic organismal depression was manifested by the depleting filtering proficiency and weakened defence against invasive Zymosan bioparticles in the phagocytosis assay. Although impaired mussels duly recovered during depuration, individuals affected by nano-PS showed immunocompetence deficiency and gill responses that were not readily reversible, which could potentially increase their vulnerability towards further environmental stressors.

Population-based variations of a core resistome revealed by urban sewage metagenome surveillance.

Sewage-based surveillance is widely employed to understand the occurrence and distribution of antimicrobial resistance (AMR) in urban community. However, there are limited studies which investigated the sewage of different sources within community. The present study used metagenomics to decipher the AMR profiles in five sources: local residence's source, animal source, migrant workers' source, clinical source , and urban wastewater treatment plant influent. A core resistome of ARGs was found across all samples, accounting for 81.4%-93.3% of the abundance of total resistome with only 17.3% diversity, irrespective of the sewage sources. Clinically relevant ARGs were identified in the core resistome across all wastewater sources. This included genes conferring resistance to beta-lactams as biomarkers of hospital sewage. The pet center wastewater showed a high abundance of genes encoding resistance to tetracycline, which is a commonly used veterinary antibiotic. The resistome profile of sewage from the migrant workers' dormitories showed a slight variation to that of the local residential population, suggesting possible differences in the human gut resistome of the foreign/migrant population, with biomarkers of genes encoding resistance to fosfomycin, fosmidomycin, kasugamycin, MLS, and polymyxin. The co-localization of ARGs and plasmid, MGEs and integrative and conjugative elements (ICEs) could explain variations in the core resistome, presumably a result of high antibiotic selection pressure. Further analysis showed a specific host-associated resistance pattern, in which core hosts mediated the core resistome profile. The core BMRGs were also co-localized with MGEs/ICEs and carried by core potential bacterial hosts. Local healthy population carried the lowest ARG load (copy number discharged by each person per day) but contributed the highest ARG burden (copy number discharged by the population). This study elucidates population-based variations of a core resistome, and further provides important insights into source tracking and management of AMR in urban environments.

Microplastics in equatorial coasts: Pollution hotspots and spatiotemporal variations associated with tropical monsoons.

Microplastics (MP < 5 mm) are eroding oceanic health and coastal development at a planetary scale. Coastlines in Southeast Asia (SEA) are plagued with plastic litters, but how MP are dispersed within SEA region is poorly understood, which can vary dramatically under the tropical climate. We systematically quantified MP in equatorial Singapore, to assess how prevailing Monsoons and other factors impact MP distributions in beaches and mangroves. Data highlighted spatial preponderance differed broadly by seasons (p < 0.05) and were strongly modulated by wind speediness (p < 0.05; r = 0.6-0.7) and promoted transboundary migrations of MP. Conversely, an inverse relationship existed between sediment MP and rainfall (r = -0.54) possibly due to re-entrainment of surficial MP. Elevated concentrations in mangrove's compartments (p < 0.05) suggest effective repository hotspots. Coastal MP consisted assorted morphologies and commonest polymers including 34% polypropylene (PP), 26% polyethelene (PE), and 23% Low Density PE. Further comparisons revealed coastal MP in Singapore accelerated by two orders of magnitude since 2014, implying cumulative pollution which is not reversible. We synthesized the first seasonal coastal MP report in SEA which is useful for source apportionment, prediction study, and mitigation planning under tropical circumstances.

Size-dependent adsorption of waterborne Benzophenone-3 on microplastics and its desorption under simulated gastrointestinal conditions.

Microplastics (MPs) are global pollutants with heightened environmental and health concerns in recent years because of their worldwide distribution across aquatic environments, ability to load chemical contaminants and the potential for ingestion by animals, including human. In this study, three commonly used and environmentally detected plastics, i.e. polystyrene, polyethylene, polypropylene with sizes of 550, 250 and 75 µm, plus two submicron-sized polystyrene microplastics (5 and 0.5 µm) were assessed as solid adsorbents for a prevalent UV filter, benzophenone-3 (BP-3). The affinity and process of adsorption exhibited differentials among different sizes and types of MPs. Apparent desorption of BP-3 from MPs under simulated gastrointestinal conditions was not significantly enhanced, which might be due to the presence of the enzyme proteins, indicating potential risk of the contaminants carried by MPs. The desorption of BP-3 from MPs was affected by the size, type of MPs and the components of the gastrointestinal fluid.

Biochar utilisation in the anaerobic digestion of food waste for the creation of a circular economy via biogas upgrading and digestate treatment.

A wood waste-derived biochar was applied to food-waste anaerobic digestion to evaluate the feasibility of its utilisation to create a circular economy. This biochar was first purposed for the upgrading of the biogas from the said anaerobic digestion, before treating and recovering the nutrients in the solid fraction of the digestate, which was finally employed as a biofertilizer for the organic cultivation of three green leafy vegetables: kale, lettuce and rocket salad. Whilst the amount of CO2 the biochar could absorb from the biogas was low (11.17 mg g-1), it could potentially be increased by modifying through physical and chemical methods. Virgin as well as CO2-laden biochar were able to remove around 31% of chemical oxygen demand, 8% of the ammonia and almost 90% of the total suspended solids from the digestate wastewater, which was better than a dewatering process via centrifugation but worse than the industry standard of a polytetrafluoroethylene membrane bioreactor. Nutrients were recovered in the solid fraction of the digestate residue filtered by the biochar, and utilised as a biofertilizer that performed similarly to a commercial complete fertilizer in terms of aerial fresh weight growth for all three vegetables cultivated. Contingent on the optimal upgrading of biogas, the concept of a circular economy based on biochar and anaerobic digestion appears to be feasible.

Modelling the spatial and seasonal distribution, fate and transport of floating plastics in tropical coastal waters.

A coupled high-resolution hydrodynamic-particle tracking model was developed to study the spatiotemporal distribution and pathways of floating plastics in the coastal waters of equatorial Singapore. The coupled model was first calibrated and validated against the field measurements and then applied to explore impact of various prevailing wind and hydrodynamic conditions on fate and transport of the plastics. The results highlighted that the wind effect on the hydrodynamics is negligible, but it influences the transmissions of floating plastics significantly in the Singapore's coastal waters. The spatial and seasonal hotspots of plastic waste were identified, which were consistent with field observations when the windage ranged from 3% to 5%. A further evaluation of the predicted trajectories showed that plastic wastes released from the land could be transported approximately 70 km seaward within 72 h when the windage was 5%. Furthermore, it was also found that the effects of climate change and increasing plastic usage would aggravate plastic pollution and accelerate its transport. The established model can provide new insights into the spatiotemporal distribution and fate of plastic waste in the tropical coastal waters, which is useful to assist regulators in making policy decisions in response to the future climate change and plastic usage.

Multidrug-resistant bacteria and microbial communities in a river estuary with fragmented suburban waste management.

River systems in developing and emerging countries are often fragmented relative to land and waste management in their catchment. The impact of inconsistent waste management and releases is a major challenge in water quality management. To examine how anthropogenic activities and estuarine effects impact water quality, we characterised water conditions, in-situ microbiomes, profiles of faecal pollution indicator, pathogenic and antibiotic resistant bacteria in the River Melayu, Southern Malaysia. Overall, upstream sampling locations were distinguished from those closer to the coastline by physicochemical parameters and bacterial communities. The abundances of bacterial DNA, total E. coli marker genes, culturable bacteria as well as antibiotic resistance ESBL-producing bacteria were elevated at upstream sampling locations especially near discharge of a wastewater oxidation pond. Furthermore, 85.7% of E. faecalis was multidrug-resistant (MDR), whereas 100% of E. cloacae, E. coli, K. pneumoniae were MDR. Overall, this work demonstrates how pollution in river estuaries does not monotonically change from inland towards the coast but varies according to local waste releases and tidal mixing. We also show that surrogate markers, such dissolved oxygen, Bacteroides and Prevotella abundances, and the rodA qPCR assay for total E. coli, can identify locations on a river that deserve immediate attention to mitigate AMR spread through improved waste management.

Impact of Redox Conditions on Antibiotic Resistance Conjugative Gene Transfer Frequency and Plasmid Fate in Wastewater Ecosystems.

Wastewater is a common pathway for the spread of antibiotic resistance (AR) genes and bacteria into the environment. Biological treatment can mitigate this path, but horizontal gene transfer (HGT) between bacteria also occurs in such processes, although the influence of bioreactor habitat and ecology on HGT frequency is not well understood. Here, we quantified how oxidation-reduction (redox) conditions impact the fate of a Green fluorescent protein (Gfp)-tagged AR plasmid (pRP4-gfp) within an E. coli host (EcoFJ1) in the liquid phase and biofilms in bioreactors. Replicate reactors treating domestic wastewater were operated under stable aerobic (+195 ± 25 mV), anoxic (-15 ± 50 mV), and anaerobic (-195 ± 15 mV) conditions, and flow cytometry and selective plating were used to quantify donor strain, EcoFJ1(pRP4-gfp), and putative transconjugants over time. Plasmid pRP4-gfp-bearing cells disappeared rapidly in aerobic ecosystems (∼2.0 log reduction after 72 h), especially in the liquid phase. In contrast, EcoFJ1(pRP4-gfp) and putative transconjugants persisted much longer in anaerobic biofilms (∼1.0 log reduction, after 72 h). Plasmid transfer frequencies were also higher under anaerobic conditions. In parallel, protozoan abundances were over 20 times higher in aerobic reactors relative to anaerobic reactors, and protozoa numbers significantly inversely correlated with pRP4-gfp signals across all reactors (p < 0.05). Taken together, observed HGT frequency and plasmid retention are impacted by habitat conditions and trophic effects, especially oxygen conditions and apparent predation. New aerobic bioreactor designs are needed, ideally employing passive aeration to save energy, to minimize resistance HGT in biological wastewater treatment processes.

Co-optimization of sponge-core bioreactors for removing total nitrogen and antibiotic resistance genes from domestic wastewater.

Inadequate sanitation can lead to the spread of infectious diseases and antimicrobial resistance (AMR) via contaminated water. Unfortunately, wastewater treatment is not universal in many developing and emerging countries, especially in rural and peri-urban locations that are remote from central sewers. As such, small-scale, more sustainable treatment options are needed, such as aerobic-Denitrifying Downflow Hanging Sponge (DDHS) bioreactors. In this study, DDHS reactors were assessed for such applications, and achieved over 79% and 84% removal of Chemical Oxygen Demand and Ammonium, respectively, and up to 71% removal of Total Nitrogen (TN) from domestic wastes. Elevated TN removals were achieved via bypassing a fraction of raw wastewater around the top layer of the DDHS system to promote denitrification. However, it was not known how this bypass impacts AMR gene (ARG) and mobile genetic element (MGE) levels in treated effluents. High-throughput qPCR was used to quantify ARG and MGE levels in DDHS bioreactors as a function of percent bypass (0, 10, 20 and 30% by volume). All systems obtained over 90% ARG reduction, although effluent ARG and TN levels differed among bypass regimes, with co-optimal reductions occurring at ~20% bypass. ARG removal paralleled bacterial removal rate, although effluent bacteria tended to have greater genetic plasticity based on higher apparent MGE levels per cell. Overall, TN removal increased and ARG removal decreased with increasing bypass, therefore co-optimization is needed in each DDHS application to achieve locally targeted TN and AMR effluent levels.

Enhanced denitrification in Downflow Hanging Sponge reactors for decentralised domestic wastewater treatment.

Enhanced aerobic/anoxic Downflow Hanging Sponge (DHS) bioreactors were assessed for carbon (C) and total nitrogen (TN) removal for decentralised domestic wastewater treatment applications. The initial design included upper aerobic and lower anoxic sponge layers, and effluent recirculation, and achieved >80% CODs and >90% NH4-N removal. However, effluent TN was higher. It was concluded the anoxic layer was C-limited for denitrification, therefore an influent bypass was added to the anoxic layer to provide supplemental C. Differed bypass ratios were compared, including 0%, 10%, 20% and 30% (% of total influent), and effluent TN declined with increasing bypass; i.e., 50.1±23.3mg-N/L, 49.9±27.8mg-N/L, 31.9±18.4mg-N/L and 10.7±5.8mg-N/L, respectively, and all reactors removed >80% CODs. This design has potential because it uses limited energy, tolerates variable flows, and simultaneously removes C and TN; all key for effective decentralised treatment applications.