Microbial communities and mobile genetic elements determine the variations of antibiotic resistance genes for a continuous year in the urban river deciphered by metagenome assembly.

Antibiotic resistance genes (ARGs) have become emerging environmental contaminants influenced by intricate regulatory factors. However, there is a lack of comprehensive studies on the evolution and distribution of ARGs over a full year in urban rivers, which serve as significant reservoirs of ARGs due to dynamic human activities. In this study, we conducted a 12-month metagenomic assembly to explore the microbial communities, ARGs, mobile genetic elements (MGEs) coexisting with ARGs, ARGs hosts, and the impact of environmental factors. Bacitracin (32%-47%) and multidrug (13%-24%) were detected throughout the year, constituting over 60% of the total abundance, making them the primary ARGs types. The assembly mechanisms of microbial communities and ARGs were primarily driven by stochastic processes. Integrase, IntI1, recombinase, and transposase were identified as the main MGEs coexisting with ARGs. Procrustes analysis revealed a significant structural association, indicating that the composition of host communities likely plays crucial roles in the seasonal composition and distribution of ARGs. Human pathogenic bacteria (HPBs) were identified in the summer, autumn, and winter, with Escherichia coli, Klebsiella pneumoniae, Acinetobacter lwoffii, and Burkholderiales bacterium being the primary HPBs. Mantle tests and PLS-PM equation analysis indicated that microbial communities and MGEs are the most critical factors determining the distribution and composition of ARGs in the river. Environmental factors (including water properties and nutrients) and ARGs hosts influence the evolution and abundance of ARGs by directly regulating microbial communities and MGEs. This study provides critical insights into risk assessment and management of ARGs in urban rivers.

Coupled effects of redox-active substances and microbial communities on reactive oxygen species in rhizosphere sediments of submerged macrophytes.

Reactive oxygen species (ROS) play crucial roles in element cycling and pollutant dynamics, but their variations and mechanisms in the rhizosphere of submerged macrophytes are poorly investigated. This study investigated the light-dark cycle fluctuations and periodic variations in ROS, redox-active substances, and microbial communities in the rhizosphere of Vallisneria natans. The results showed sustained production and significant diurnal fluctuations in the O2•- and •OH from 27.6 ± 3.7 to 61.7 ± 3.0 µmol/kg FW and 131.0 ± 6.8 to 195.4 ± 8.7 µmol/kg FW, respectively, which simultaneously fluctuated with the redox-active substances. The ROS contents in the rhizosphere were higher than those observed in non-rhizosphere sediments over the V. natans growth period, exhibiting increasing-decreasing trends. According to the redundancy analysis results, water-soluble phenols, fungi, and bacteria were the main factors influencing ROS production in the rhizosphere, showing contribution rates of 74.0, 17.3, and 4.4 %, respectively. The results of partial least squares path modeling highlighted the coupled effects of redox-active substances and microbial metabolism. Our findings also demonstrated the degradation effect of ROS in rhizosphere sediments of submerged macrophytes. This study provides experimental evidence of ROS-related rhizosphere effects and further insights into submerged macrophytes-based ecological restoration.

The coupling effect promotes superoxide radical production in the microalgal-fungal symbiosis systems: Production, mechanisms and implication for Hg(II) reduction.

Redox transformation of mercury (Hg) is critical for Hg exchange at the air-water interface. However, the superoxide radicals (O2•─) contribution of microalgal-fungal symbiotic systems in lake water to Hg(II) reduction is mainly unknown. Here, we studied the enhanced potential for O2•─ production by the coupling effect between microalgae and fungi. The relationships between microenvironment, microorganisms, and O2•─ production were also investigated. Furthermore, the implication of O2•─ for Hg(II) reduction was explored. The results showed that the coupling effect of microalgae and fungi enhanced O2•─ generation in the symbiotic systems, and the O2•─ generation peaked on day 4 in the lake water at 160.51 ± 13.06-173.28 ± 18.21 µmol/kg FW (fresh weight). In addition, O2•- exhibited circadian fluctuations that correlated with changes in dissolved oxygen content and redox potential on the inter-spherical interface of microalgal-fungal consortia. Partial least squares path modeling (PLS-PM) indicates that O2•─ formation was primarily associated with microenvironmental factors and microbial metabolic processes. The experimental results suggest that O2•─ in the microalgal-fungal systems could mediate Hg(II) reduction, promoting Hg conversion and cycling. The findings highlight the importance of microalgae and fungal symbiotic systems in Hg transformation in aquatic environments.

Evolution and distribution of antibiotic resistance genes in submerged macrophytes and biofilm systems: From seasonal monitoring to mesocosm experiments.

The aquatic ecosystem has been extensively investigated as a hotspot for the spread of antibiotic resistance genes (ARGs); yet, the evolution and distribution of ARGs profiles in submerged macrophytes biofilms and surrounding water remained unclear. In this study, the dynamic distribution and seasonal variations of microbial communities and ARGs profiles were investigated, alongside their assembly processes and mutual interactions. Bacitracin and multidrug resistance genes were predominant, constituting more than 60% of the total ARGs abundance. The deterministic processes (<65%), influenced by the physicochemical properties of the river environment, governed the assembly and composition of ARGs profiles, exhibiting significant seasonal variation. The peak diversity (21 types) and abundance (0.316 copy ratios) of ARGs were detected during the summer. Proteobacteria and Actinobacteria were the dominant bacterial phyla, accounting for 38.41-85.50% and 4.03-27.09% of the microbial community, respectively. Furthermore, Proteobacteria, especially genera such as Acinetobacter, Burkholderia, and Pseudomonas, with various resistance sequences, were the primary carriers of multiple ARGs. Notably, the genetic exchanges between biofilms and surrounding water facilitated the further propagation of high-risk ARGs, posing greater ecological risks. Redundancy analysis indicated that the total nitrogen and temperature in water determined the fate of pathogenic-resistant species. These findings provided theoretical support for the mitigation of ARGs contamination in aquatic environments.

Purification of aquaculture wastewater by macrophytes and biofilm systems: Efficient removal of trace antibiotics and enrichment of antibiotic resistance genes.

The purification performance of aquaculture wastewater and the risk of antibiotic resistance genes (ARGs) dissemination in wetlands dominated by macrophytes remain unclear. Here, the purification effects of different macrophytes and biofilm systems on real aquaculture wastewater were investigated, as well as the distribution and abundance of ARGs. Compared to the submerged macrophytes, artificial macrophytes exhibited higher removal rates of TOC (58.80 ± 5.04 %), TN (74.50 ± 2.50 %), and TP (77.33 ± 11.66 %), and achieved approximately 79.92 % removal of accumulated trace antibiotics in the surrounding water. Additionally, the biofilm microbial communities on the surface of artificial macrophytes exhibited higher microbial diversity with fewer antibiotic-resistant bacteria (ARB) enrichment from the surrounding water. The absolute abundance of ARGs (sul1, sul2, and intI1) in the mature biofilm to be one to two orders of magnitude higher than that in the water. Although biofilms could decrease ARGs in the surrounding water by enriching ARB, the intricate network structure of biofilms further facilitated the proliferation of ARB and the dissemination of ARGs in water. Network analysis suggested that Proteobacteria and Firmicutes phyla were dominant and potential carriers of ARGs, contributing 69.00 % and 16.70 %, respectively. Our findings highlight that macrophytes and biofilm systems have great performance on aquaculture wastewater purification, but with high risk of ARGs.

Effect of different size microplastic particles on the construction of algal-bacterial biofilms and microbial communities.

Algal-bacterial symbiotic system is a biological purification system that combines sewage treatment with resource utilization and has the dual effects of carbon sequestration and pollution reduction. In this study, an immobilized algal-bacterial biofilm system was constructed for the treatment of natural sewage. Effects of exposure to microplastics (MPs) with different particle diameters (0.065 µm, 0.5 µm and 5 µm) were determined in terms of algal biomass recovery efficiency, the composition of extracellular polymeric substances (EPS) and morphologic characteristics. The impacts of MPs on the bacterial diversity and community structure of biofilms were also examined. The metagenomic analysis of key microorganisms and related metabolism pathways involved in system was further investigated. Results showed that following exposure to 5 µm MP, a maximum algal recovery efficiency of 80% was achieved, with a minimum PSII primary light energy conversion efficiency (Fv/Fm ratio) of 0.513. Furthermore, 5 µm MP caused the highest level of damage to the algal-bacterial biofilm, enhancing the secretion of protein-rich EPS. The biofilm morphology became rough and loose following exposure to 0.5 µm and 5 µm MP. Community diversity and richness were significantly high in biofilms exposed to 5 µm MP. Proteobacteria (15.3-24.1%), Firmicutes (5.0-7.8%) and Actinobacteria (4.2-4.9%) were dominant in all groups, with exposure to 5 µm MP resulting in the highest relative abundance for these species. The addition of MPs promoted the related metabolic functions while inhibited the degradation of harmful substances by algal-bacterial biofilms. The findings have environmental significance for the practical application of algal-bacterial biofilms for sewage treatment, providing novel insights into the potential effects of MPs on immobilized algal-bacterial biofilm systems.

Seasonal changes driving shifts of aquatic rhizosphere microbial community structure and the functional properties.

Ecological floating beds could enable roots to become suspended and this allowed submerged roots to harbour various types of microbes. But, there was a lack of systematic research on microbial community structure changes and the influencing mechanisms. In this study, the ecological floating beds were constructed using selected plants [Cyperus involucratus Rottboll (Cyp), Thalia dealbata Fraser (Tha) and Iris tectorum Maxim (Iri)] that was compared with a control group [static water (S)]. The results showed that the highest abundance and diversity of root microbial communities were found in autumn, with the dominant taxa being Proteobacteria, Actinobacteriota, Cyanobacteria, Chloroflexi, Firmicutes, Bacteroidota, and Acidobacteriota. The microbial communities of Tha and Cyp groups greatly overlapped, while the Iri and control groups exhibited distinctly diverse communities. The root microbial populations of the same plant also reflected a large change in different seasons. Conversely, photosynthetic autotrophs and specialized anaerobes were more inclined to thrive at higher temperatures and lower DO concentrations and then they gradually became the dominant species. Microbial co-occurrences of the Tha and control groups were complex and showed both cooperation and competition. In addition, TOC was an important environmental factor that shaped the microbial community structures and DO changed the microbial community by affecting the abundance of aerobic and anaerobic bacteria. Microorganisms showed potential for degradation and metabolism of non-food substances with low/no corresponding metabolic pathways.

Response of submerged macrophytes and leaf biofilms to different concentrations of oxytetracycline and sulfadiazine.

Oxytetracycline and sulfadiazine were widely used and they entered the environment through various channels such as domestic sewage, medical wastewater and agricultural wastewater, causing significant ecological risk. To determine the effects of different antibiotic concentrations on submerged macrophytes, Vallisneria natans was exposed to solutions containing different concentrations of oxytetracycline and sulfadiazine (0.1 mg/L、1 mg/L、10 mg/L、50 mg/L). After 20-days exposure, we found that 10 mg/L groups had a significant effect on Vallisneria natans. Under high antibiotic concentrations, the growth of Vallisneria natans was inhibited, chloroplasts were deformed, the chlorophyll content was reduced, and antioxidant enzyme activities, such as superoxide dismutase and glutathione, were increased. There was no significant difference between the control group and groups with low antibiotic concentrations (≤1 mg/L). The N-acyl-l-homoserine lactone concentration tended to increase with increasing antibiotic concentrations. The presence of antibiotics also affected the microbial community structure of biofilms on the submerged macrophytes. For example, the higher the concentration of antibiotics, the higher the proportion of Proteobacteria. These results suggest that high concentrations of oxytetracycline and sulfadiazine can disrupt homeostasis, induce effective Vallisneria natans defense mechanisms and alter biofilms in aquatic ecosystems.