International audience | The reuse of treated wastewater (TWW) for irrigation is widely applied to alleviate pressure on freshwater resources. However, TWW contains antibiotics that once in soils, can exert selective pressure, promoting the emergence and spread of antimicrobial resistance (AMR) in the environment. Current environmental risk assessments for antibiotic residues rely on indicators such as Predicted No Effect Concentrations (PNECs), usually determined in liquid media. These PNECs aim to predict antibiotic concentrations that may promote resistance in the environment. Given the complexity of soil matrices, few studies have established PNEC values for soil, which likely differ significantly from aquatic environments.To address this gap, we developed a simplified experimental model using soil microcosms irrigated with TWW and the antibiotic sulfamethoxazole (SMX) to estimate threshold concentrations favouring resistance transfer or/and emergence within the soil microbiome. We identified SMX concentrations between 0.01 and 0.1 mg/kgdry soil that likely increased the abundance of sulfonamide resistance genes in soil. A time window of 1 to 7 days post-exposure showed a temporary rise in sul1 and intl1 gene abundance (over 1 log/soil 16S rDNA), the appearance of SMX transformation products, and an increase in some Rhodocyclaceae. After 1.5 months of incubation and complete SMX transformation, the relative abundance of sul1 and intl1 remained about 0.5 log higher than in SMX-free controls and soils with SMX levels below 0.1 mg/kg dry soil. A persistent transformation product, 4-N-glucuronide-SMX, was also observed.Here, the estimated PNEC for SMX in soil, between 0.01 and 0.1 mg/kg, exceeds typical SMX concentrations found in soils exposed to TWW. This may suggest low impact on resistance selection for this compound in the context of TWW exposure. However further studies on other soils, water, and antibiotics need to be conducted to expand our knowledge on soil PNECs.

Antibiotics are a contaminant class of worldwide concern as they are frequently detected in aquatic ecosystems. To better understand the impacts of antibiotics on aquatic ecosystems, we conducted an outdoor mesocosm experiment in which aquatic communities were exposed to different concentrations of the antibiotic sulfamethoxazole (0, 0.15, 1.5, 15 and 150 μg/L). These concentrations include mean (0.15 μg/L) and maximum detected concentrations (15 and 150 μg/L) in aquatic ecosystems worldwide. Sulfamethoxazole was applied once a week for eight consecutive weeks to 1530 L outdoor mesocosms in the Netherlands, followed by an eight-week recovery period. We evaluated phytoplankton-, bacterial- and invertebrate responses during and after sulfamethoxazole exposure and assessed impacts on organic matter decomposition. Contrary to our expectations, consistent treatment-related effects on algal and bacterial communities could not be demonstrated. In addition, sulfamethoxazole did not significantly affect zooplankton and macroinvertebrate communities. However, some effects on specific taxa were observed, with an increase in Mesostoma flatworm abundance (NOEC of <0.15 μg/L). In addition, eDNA analyses indicated negative impacts on the insects Odonata at a sulfamethoxazole concentration of 15 μg/L. Overall, environmentally relevant sulfamethoxazole concentration did not result in direct or indirect impairment of entire aquatic communities and ecological processes in our mesocosms. However, several specific macroinvertebrate taxa demonstrated significant (in)direct effects from sulfamethoxazole. Comparison of the results with the literature showed inconsistent results between studies using comparable, environmentally relevant, concentrations. Therefore, our study highlights the importance of testing the ecological impacts of pharmaceuticals (such as sulfamethoxazole) across multiple trophic levels spanning multiple aquatic communities, to fully understand its potential ecological threats.

Emission of antibiotics into riverine environments affects aquatic ecosystem functions and leads to the development of antibiotic resistance. Here, the profiles of forty-four antibiotics and eighteen antibiotic resistance genes (ARGs) were analyzed in two large rivers of the Pearl River System. In addition, the risks of ecotoxicity and resistance selection posed by the antibiotics were estimated. As compared to the reservoirs, the river sections close to the urban and livestock areas contained more antibiotics and ARGs. Seasonal variations of antibiotics (higher in the dry season) and relative ARGs (normalized by 16S rRNA gene, higher in the wet season) were found in the water, but not in the sediment. Sulfonamide resistance genes were the most prevalent ARGs in both river water and sediment. Antibiotic concentration was correlated with ARG abundance in the water, indicating that antibiotics play a critical role in ARG spread. In addition, oxytetracycline was the most abundant antibiotic with concentrations up to 2030 ng/L in the water and 2100 ng/g in the sediment respectively, and posed the highest risks for resistance selection. Oxytetracycline, tetracycline and sulfamethoxazole were expected to be more ecotoxicologically harmful to aquatic organisms, while ofloxacin, enrofloxacin, norfloxacin, chlortetracycline, oxytetracycline and tetracycline posed ecotoxicological risks in the sediment. The Nanliujiang river with intensive livestock activities was contaminated by antibiotics and ARGs and faced high ecotoxicological and resistance selection risks. Collectively, these findings reflect the impacts of anthropogenic activities on the spread of antibiotic resistance in large river basins.

Aquatic pollution from emerging organic contaminants (EOCs) is of key environmental importance in India and globally, particularly due to concerns of antimicrobial resistance, ecotoxicity and drinking water supply vulnerability. Here, using a broad screening approach, we characterize the composition and distribution of EOCs in groundwater in the Gangetic Plain around Patna (Bihar), as an exemplar of a rapidly developing urban area in northern India. A total of 73 EOCs were detected in 51 samples, typically at ng.L⁻¹ to low μg.L⁻¹ concentrations, relating to medical and veterinary, agrochemical, industrial and lifestyle usage. Concentrations were often dominated by the lifestyle chemical and artificial sweetener sucralose. Seventeen identified EOCs are flagged as priority compounds by the European Commission, World Health Organisation and/or World Organisation for Animal Health: namely, herbicides diuron and atrazine; insecticides imidacloprid, thiamethoxam, clothianidin and acetamiprid; the surfactant perfluorooctane sulfonate (and related perfluorobutane sulfonate, perfluorohexane sulfonate, perfluorooctanoic acid and perfluoropentane sulfonate); and medical/veterinary compounds sulfamethoxazole, sulfanilamide, dapson, sulfathiazole, sulfamethazine and diclofenac. The spatial distribution of EOCs varies widely, with concentrations declining with depth, consistent with a strong dominant vertical flow control. Groundwater EOC concentrations in Patna were found to peak within ∼10 km distance from the River Ganges, indicating mainly urban inputs with some local pollution hotspots. A heterogeneous relationship between EOCs and population density likely reflects confounding factors including varying input types and controls (e.g. spatial, temporal), wastewater treatment infrastructure and groundwater abstraction. Strong seasonal agreement in EOC concentrations was observed. Co-existence of limited transformation products with associated parent compounds indicate active microbial degradation processes. This study characterizes key controls on the distribution of groundwater EOCs across the urban to rural transition near Patna, as a rapidly developing Indian city, and contributes to the wider understanding of the vulnerability of shallow groundwater to surface-derived contamination in similar environments.

The negative impact of antibiotic pollution on the agricultural system and human health is a hot issue in the world. However, little information is available on the antibiotics toxicity mechanism and the role of silicon (Si) to alleviate the antibiotics toxicity. In this study, strawberry (Fragaria ananassa) showed excitatory response to low-dose SMZ (1 mg L⁻¹), but strawberry root and photosynthetic efficiency were damaged under high level. When SMZ level exceeded 10 mg L⁻¹, H₂0₂, O₂⁻, MDA and relative conductivity increased, while SOD and CAT activities first increased and then decreased. SMZ accumulated more in roots and fruits, but less in stems, and the accumulation increased with the increase of SMZ-dose. Under 1 mg L⁻¹ SMZ, the SMZ accumulation in fruits was 110.54 μg kg⁻¹, which exceeded the maximum residue limit. SMZ can induce the expression of sul1, sul2 and intI1, and intI1 had the highest abundance. Exogenous application of Si alleviated the toxicity of SMZ, which is mainly related to the degradation of SMZ in soil and the reduction of SMZ absorption by strawberry. In addition, Si relieved root damage, promoted the increase of photosynthetic efficiency, and improved the antioxidant system to resist SMZ toxicity.

Antibiotics can stimulate the growth of model cyanobacterial species under pure culture conditions, but their influence on cyanobacterial blooms in natural aquatic ecosystems remains unclear. In this study, three commonly detected antibiotics (sulfamethoxazole, tetracycline, and ciprofloxacin) and their ternary mixture were proved to selectively stimulate (p < 0.05) the growth and photosynthetic activity of cyanobacteria in an aquatic microcosm at an environmentally relevant exposure dose of 300 ng/L under both oligotrophic and eutrophic conditions. Under the eutrophic condition, cyanobacteria reached a bloom density of 1.61 × 10⁶ cells/mL in 15 days without antibiotics, while the cyanobacteria exposed to tetracycline, sulfamethoxazole, ciprofloxacin, and their ternary mixture exceeded this bloom density within only 10, 8, 7, and 6 days, respectively. Principal coordinate analysis indicated that the antibiotic contaminants accelerated the prokaryotic community succession towards the formation of a cyanobacterial bloom by promoting the dominance of Microcystis, Synechococcus, and Oscillatoria under the eutrophic condition. After 15 days of culture, the antibiotic exposure increased the density of cyanobacteria by 1.38–2.31-fold and 2.28–3.94-fold under eutrophic and oligotrophic conditions, respectively. Antibiotic exposure generated higher stimulatory effects on cyanobacterial growth under the oligotrophic condition, but the antibiotic(s)-treated cyanobacteria did not form a bloom due to nutrient limitation. Redundancy analysis indicated that the three target antibiotics and their ternary mixture affected the prokaryotic community structure in a similar manner, while tetracycline showed some differences compared to sulfamethoxazole, ciprofloxacin, and the ternary antibiotic mixture with regard to the regulation of the eukaryotic community structure. This study demonstrates that antibiotic contaminants accelerate the formation of cyanobacterial blooms in eutrophic lake water and provides insights into the ecological effects of antibiotics on aquatic microbial communities.

Traditional composting has already shown a certain effect in eliminating antibiotic residues, antibiotic-resistant bacteria (ARBs), and antibiotic resistance genes (ARGs). It is worth noting that the rebounding of ARGs and the succession of the bacterial community during conventional aerobic composting are still serious threats. Considering the probable risk, improved and adaptable technologies are urgently needed to control antibiotic resistance efficiently. This study monitored how thermophilic aerobic composting affected the ARGs, as well as the bacterial diversity during the composting of cow manure spiked with sulfamethoxazole (SMX) at different concentrations. Results showed that the degradation of SMX was enhanced during thermophilic aerobic composting (control > SMX25 > SMX50 > SMX100) and was no longer detected after 20 days of composting. High temperature or heat significantly stimulated the rebounding of certain genes. After 35 days, the abundance of detected genes (sul2, sulA, dfrA7, and dfrA1) significantly decreased (p < 0.05) in control and antibiotic-spiked treatments, except for sul1. The addition of three concentrations of SMX elicited a sharp effect on bacterial diversity, and microbial structure in SMX25 led to significant differences with others (p < 0.05). The network analysis revealed more rigorous interactions among ARGs and abundant genera, suggesting that the host of ARGs potentially increased at low concentrations of SMX. Especially, genera g_norank_f__Beggiatoaceae, Ruminiclostridium, Caldicoprobacter, g_norank_o_MBA03, Hydrogenispora, and Ruminiclostridium_1 were major potential hosts for sul1. In conclusion, the rebounding of ARGs could be intermitted partially, and more efficient control of antibiotic resistance could be achieved in the thermophilic composting compared to conventional methods.

The pollution of wastewater with antibiotics and antibiotics resistance genes has attracted public concerns about ecosystem and global health. Swine wastewater can contain high concentrations of antibiotics, especially sulfonamides, even after full-scale wastewater treatment. In this study, mesocosm-scale vertical flow constructed wetlands (VF-CWs) were applied to abate nutrients and antibiotics in swine wastewater containing sulfonamides. VF-CWs performed well in the removal of both nutrients and antibiotics. Sulfonamides did not influence total organic carbon (TOC) and total phosphorus (TP) removal, and even slightly enhanced NH₄⁺–N removal. High removal efficiencies (26.42–84.05%) were achieved for sulfadiazine (SDZ), sulfamethoxazole (SMX) and sulfamethazine (SMZ). Together with lab-scale sorption and biodegradation experiments, microbial degradation was found to be the most important removal mechanism for sulfonamides in VF-CWs. Sulfonamides addition increased bacterial alpha-diversity and changed microbial community structure. Moreover, antibiotics promoted antibiotic-resistant or -degrading bacteria. Bacillus, Geobacter and other seven genera were correlated with sulfonamides reduction under either aerobic or anaerobic condition. In summary, VF-CW is a suitable alternative for swine wastewater treatment, and biodegradation plays the key role in sulfonamides abatement.Main findings of the work.This was the first work to combine bacterial community analysis with microcosm experiments to uncover the major removal mechanism of sulfonamides in constructed wetlands.

The potential risk to human health of antibiotics that pass through the food chain has become an important global issue, but there are few reports on the response of ginger (Zingiber officinale) to antibiotic pollution. In this study, we investigated the enrichment characteristics and biological response of ginger to sulfamethoxazole (SMZ) and ofloxacin (OFL) residues, which are common in the environment. Lower levels of SMZ, OFL and their combined duplex treatment (SMZ+OFL) promoted the growth of ginger, but the critical doses necessary to stimulate growth differed among treatments: 10 mg L⁻¹ SMZ, 1 mg L⁻¹ OFL and 1 mg L⁻¹ (SMZ+OFL) had the strongest stimulating effects. At higher dosages, the root growth and light energy utilization efficiency of ginger were impaired, and (SMZ+OFL) had the strongest inhibitory effect. Treatments with lower levels of antibiotics had no significant effect on reactive oxygen species and antioxidant enzyme activities. However, when SMZ, OFL and SMZ+OFL concentrations exceeded 10 mg L⁻¹, the contents of H₂O₂, O₂⁻ and MDA continued to increase, while the activities of SOD, POD, CAT first increased and then decreased, especially in SMZ+OFL. Ginger accumulated more SMZ and OFL in rhizomes and less in leaves, and accumulation increased significantly as antibiotic concentration increased. When SMZ concentration was 1 mg L⁻¹, the SMZ concentrations in rhizomes, roots, and leaves were 0.23, 0.15, and 0.05 mg kg⁻¹, respectively, and the residual SMZ in the rhizome was 2.3 times higher than the maximum residue limit. The abundance of the resistance genes sul1, sul2, qnrS, and intI1 increased with increasing antibiotic concentrations, and intI1 abundance was the highest. OFL induced higher levels of intI1 expression than did SMZ.

Recently, the widespread use of antibiotic has raised concerns about the potential health risks associated with their microbiological effect. In the present study, we investigated 990 elderly individuals (age ≥ 60 years) from the Cohort of Elderly Health and Environment Controllable Factors in West Anhui, China. A total of 45 representative antibiotics and two antibiotic metabolites were monitored in urine samples through liquid chromatography electrospray tandem mass spectrometry. The results revealed that 34 antibiotics were detected in 93.0% of all urine samples and the detection frequencies of each antibiotic varied between 0.2% and 35.5%. The overall detection frequencies of seven human antibiotics (HAs), 10 veterinary antibiotics (VAs), three antibiotics preferred as HAs (PHAs), and 14 preferred as VAs (PVAs) in urines were 27.4%, 62.9%, 30.9% and 72.7%, respectively. Notably, the samples with concentrations of six PVAs (sulfamethoxazole, trimethoprim, oxytetracycline, danofloxacin, norfloxacin and lincomycin) above 5000 ng/mL accounted for 1.7% of all urine samples. Additionally, in 62.7% of urine samples, the total antibiotic concentration was in the range of the limits of detection to 20.0 ng/mL. Furthermore, the elderly individuals with the sum of estimated daily intakes of VAs and PVAs more than 1 μg/kg/day accounted for 15.2% of all participants, and a health risk related to change in gut microbiota under antibiotic stimulation was expected in 6.7% of the elderly individuals. Especially, ciprofloxacin was the foremost contributor to the health risk, and its hazard quotient value was more than one in 3.5% of all subjects. Taken together, the elderly Chinese people were extensively exposed to VAs, and some elderly individuals may have a health risk associated with dysbiosis of the gut microbiota.