Metagenomic profiling of antibiotic resistance genes/bacteria removal in urban water: Algal-bacterial consortium treatment system.

Antibiotic resistance genes (ARGs) have exhibited significant ecological concerns, especially in the urban water that are closely associated with human health. In this study, with presence of exogenous Chlorella vulgaris-Bacillus licheniformis consortium, most of the typical ARGs and MGEs were removed. Furthermore, the relative abundance of potential ARGs hosts has generally decreased by 1-4 orders of magnitude, revealing the role of algal-bacterial consortium in cutting the spread of ARGs in urban water. While some of ARGs such as macB increased, which may be due to the negative impact of algicidal bacteria and algal viruses in urban water on exogenous C. vulgaris and the suppression of exogenous B. licheniformis by indigenous microorganisms. A new algal-bacterial interaction might form between C. vulgaris and indigenous microorganisms. The interplay between C. vulgaris and bacteria has a significant impact on the fate of ARGs removal in urban water.

Toxicity effects of disinfection byproduct chloroacetic acid to Microcystis aeruginosa: Cytotoxicity and mechanisms.

Chlorine-based disinfectants are widely used for disinfection in wastewater treatment. The mechanism of the effects of chlorinated disinfection by-products on cyanobacteria was unclear. Herein, the physiological effects of chloroacetic acid (CAA) on Microcystis aeruginosa (M. aeruginosa), including acute toxicity, oxidative stress, apoptosis, production of microcystin-LR (MC-LR), and the microcystin transportation-related gene mcyH transcript abundance have been investigated. CAA exposure resulted in a significant change in the cell ultrastructure, including thylakoid damage, disappearance of nucleoid, production of gas vacuoles, increase in starch granule, accumulation of lipid droplets, and disruption of cytoplasm membranes. Meanwhile, the apoptosis rate of M. aeruginosa increased with CAA concentration. The production of MC-LR was affected by CAA, and the transcript abundance of mcyH decreased. Our results suggested that CAA poses acute toxicity to M. aeruginosa, and it could cause oxidative damage, stimulate MC-LR production, and damage cell ultrastructure. This study may provide information about the minimum concentration of CAA in the water environment, which is safe for aquatic organisms, especially during the global coronavirus disease 2019 pandemic period.

Removal mechanism of Microcystis aeruginosa in Fe2+/sodium percarbonate and Fe2+/sodium persulfate advanced oxidation-flocculation system.

Advanced oxidation process (AOPs) can be used for the treatment of harmful algal blooms (HABs). In this study, two systems of Fe2+/sodium percarbonate (Fe2+/SPC system) and Fe2+/sodium persulfate (Fe2+/PS system) were established to explore the removal mechanism of Microcystis aeruginosa (M. aeruginosa). The results indicated that the Fe2+/SPC system catalyzed H2O2 to generate a large amount of [Formula: see text] for oxidation by Fe2+ and formed Fe3+ to promote the flocculation of M. aeruginosa. The persulfate was activated by Fe2+ to generate [Formula: see text] with super-oxidizing properties, and Fe3+ was generated to realize the oxidation and flocculation of M. aeruginosa in the Fe2+/PS system. Compared with the traditional method in which the pre-oxidation and flocculation processes are carried out separately, the method in this study effectively improves the utilization rate of the flocculant and the removal effect of M. aeruginosa. The absolute value of zeta potential of Fe2+/PS system (|ζ|= 0.808 mV) was significantly lower than that of Fe2+/SPC system (|ζ|= 21.4 mV) (P < 0.05), which indicated that Fe2+/PS system was more favorable for the flocculation of M. aeruginosa cells than the Fe2+/SPC system.

Algal-bacterial consortium mediated system offers effective removal of nitrogen nutrients and antibiotic resistance genes.

The sulfonamide antibiotic resistance genes (ARGs) especially sul1 was identified as the dominant in eutrophic water. The performance of Chlorella vulgaris-B. licheniformis consortium toward sul1 removal, total nitrogen (TN) removal, and the mechanism of sul1 removal was investigated. The removal efficiency of exogenous ARGs plasmids carrying sul1 reached (97.2 ± 2.3)%. The TN removal rate reached (98.5 ± 1.2)%. The enhancements of carbon metabolism, nitrogen metabolism, aminoacyl-tRNA biosynthesis, and glycoproteins had significant influences on sul1 and TN removals, under the premise of normal growth of algae and bacteria. The quantitative polymerase chain reaction (qPCR) results suggested that the absolute abundances of sul1 were low in algal-bacterial systems (0 gene copies/mL) compared with individual systems ((1 × 106 ± 15) gene copies/mL). The duplication of sul1 was inhibited in algal cells and bacterial cells. The algal-bacterial consortium seems to be a promising technology for wastewater treatment with a potential to overcome the eutrophication and ARGs challenges.

Enantioselective effects of chiral fragrance carvone (L- and D-carvone) on the physiology, oxidative damage, synthesis, and release of microcystin-LR in Microcystis aeruginosa.

Carvone is a widely used chiral fragrance with two isomers (L-carvone and D-carvone). D-carvone smells like a caraway, whereas L-carvone smells like mint. Carvone imposes a potential burden on the aquatic ecosystem. However, the enantioselective toxic effect of carvone enantiomers on cyanobacteria remains unknown. This study aims to investigate the effects of L- and D-carvone on the physiological processes and related gene transcription (phoU, rbcL, and mcyH) in M. aeruginosa. Results showed that in the presence of L- and D-carvone, the oxidative damage and inhibitory effects on growth occurred in a concentration-dependent manner. The contents of chlorophyll a and protein and the rbcL transcription level were inhibited in M. aeruginosa. In addition, intracellular adenosine triphosphate (ATP) was heavily depleted because of various biological processes, including growth, oxidation reactions, and gene regulation. Meanwhile, L- and D-carvone stimulated the production and release of MC-LR and upregulated the expression level of the MC-LR-related gene mcyH. Intracellular MC-LR likely leaked to the water body under L-carvone exposure, posing a potential threat to the water environment. This study indicated that L- and D-carvone can regulate the physiological and metabolic activity of M. aeruginosa and show enantioselective toxic effects. The findings will also provide important insights into the influence of chiral fragrance on cyanobacterial blooms. Furthermore, this study will guide the safe application of chiral fragrance as personal care products.

Mechanism of enhanced production of triterpenoids in algal-fungal consortium.

Chlorella pyrenoidosa-Ganoderma lucidum symbiotic systems were constructed. The mechanism of enhanced production of triterpenoids in algal-fungal consortium by comparing the contents of triterpenoids in individual fungal systems and algal-fungal consortium systems was investigated. The production of triterpenoids in C. pyrenoidosa-G. lucidum consortium increased significantly (P < 0.05). The categories and relative abundances of metabolites in the individual systems and algal-fungal systems were measured and analyzed by metabonomic tests. There were 57 significant different metabolites (VIP > 1 and P < 0.05) including 12 downregulated metabolites and 45 upregulated metabolites were obtained. The significant enriched metabolic pathways (VIP > 1 and P < 0.05) of citrate cycle (TCA cycle), tyrosine metabolism, glycolysis, and terpenoid backbone biosynthesis in algal-fungal consortium were obtained. The relative abundances of important precursors of triterpenoids including mevalonic acid, lanosterol, and hydroquinone were 1.4 times, 1.7 times, and 2 times, respectively, in algal-fungal consortium than that in the individual fungal systems. The presence of C. pyrenoidosa in algal-fungal consortium promoted the biosynthesis of triterpenoids in G. lucidum.

Research progress of advanced oxidation technology for the removal of Microcystis aeruginosa: a review.

In recent years, cyanobacteria blooms have continued to erupt frequently, seriously jeopardizing the safety of drinking water and human health. The safe, quick, and economical removal of cyanobacteria from water bodies, especially the dominant species of cyanobacteria, Microcystis aeruginosa, has captured a lot of scientists' attention. The application of advanced oxidation technology in water treatment is very promising, but it has not yet been used in production. To further promote the application of the advanced oxidation method in water treatment, this article combines the results of advanced research in China and abroad to review this emergent technology. Briefly, advanced oxidation process methods employ various mechanisms to remove the dominant species of cyanobacteria blooms Microcystis aeruginosa. This provides a theoretical reference and support for the efficient removal of harmful cyanobacteria from water.

Mutual impacts and interactions of antibiotic resistance genes, microcystin synthetase genes, graphene oxide, and Microcystis aeruginosa in synthetic wastewater.

The physiological impacts and interactions of antibiotic resistance gene (ARG) abundance, microcystin synthetase gene expression, graphene oxide (GO), and Microcystis aeruginosa in synthetic wastewater were investigated. The results demonstrated that the absolute abundance of sul1, sul2, tetW, and tetM in synthetic wastewater dramatically increased to 365.2%, 427.1%, 375.2%, and 231.7%, respectively, when the GO concentration was 0.01 mg/L. Even more interesting is that the sum gene copy numbers of mcyA-J also increased to 243.2%. The appearance of GO made the significant correlation exist between ARGs abundance and mcyA-J expression. Furthermore, M. aeruginosa displayed better photosynthetic performance and more MCs production at 0.01 mg/L GO. There were 65 pairs of positive correlations between the intracellular differential metabolites of M. aeruginosa and the abundance of sul1, sul2, tetM, and tetW with various GO concentrations. The GO will impact the metabolites and metabolic pathway in M. aeruginosa. The metabolic changes impacted the ARGs, microcystin synthetase genes, and physiological characters in algal cells. Furthermore, there were complex correlations among sul1, sul2, tetM, tetW, mcyA-J, MCs, photosynthetic performance parameters, and ROS. The different concentration of GO will aggravate the hazards of M. aeruginosa by promoting the expression of mcyA-J, producing more MCs; simultaneously, it may cause the spread of ARGs.

The effect of carbon-based copper nanocomposites on Microcystis aeruginosa and the movability of antibiotic resistance genes in urban water.

The presence of Microcystis aeruginosa (M. aeruginosa) can affect the transference of antibiotic resistance genes (ARGs), and the presence of carbon-based copper nanocomposites (CCN) can affect the growth of M. aeruginosa. However, the effect of CCN on M. aeruginosa and ARGs is not fully understood. In this study, metagenomic sequencing was employed to analyze the movability of ARGs, their potential transfer, and possible hosts in photobioreactor treating urban water. The results uggested that 20 mg/L of CCN changed the composition and abundance of microorganisms in urban water, significantly promoted the flocculation of M aeruginosa, and decreased the composing proportion of Cyanophyta sp. and M aeruginosa. The results indicated that 20 mg/L of CCN significantly decreased the absolute abundance and ARGs proportions which mediated by plasmids (32.7 %). Furthermore, the lower co-occurrence probability of ARGs and mobile genetic elements (MGEs) suggested that 20 mg/L of CCN weakened the movability potential of ARGs mediated by MGEs such as plasmids. Among the 452 metagenome-assembled genomes (MAGs), 95 MAGs belonging to 41 bacterial categories were identified as possible ARG hosts. These results will provide insights into the control of harmful cyanobacteria and the management of ARGs in urban water.

Effects of exogenous vitamin B12 on nutrient removal and protein expression of algal-bacterial consortium.

Chlorella vulgaris and Bacillus licheniformis consortium was added to synthetic wastewater with exogenous vitamin B12. In the presence of 100 ng/L vitamin B12, removal efficiencies of TN, NH3-N, PO43-P, and COD were 80.1%, 76.8%, 87.9%, and 76.7%, respectively. The functional groups on the cell surface of the consortium, including -NH, -CH3, C=O, C=C, and P-O-C, increased with 100 ng/L vitamin B12. These functional groups improved the biological adsorption of the consortium; however, higher concentrations of vitamin B12 resulted in an occlusion of the functional groups. Furthermore, there were 5 significantly enriched protein pathways, namely carbon fixation in photosynthetic organisms; amino acid metabolic pathways; the pathway of one carbon pool by folate; nitrogen metabolism; and photosynthesis. Most proteins in these pathways were upregulated, which enhanced carbon fixation and photosynthesis in the algal cells. Simultaneously, B12 promoted significant upregulation of proteins associated with the quorum-sensing pathway, which promoted the interaction between algae and bacteria.

Effects of graphene oxide on algal cellular stress response: Evaluating metabolic characters of carbon fixation and nutrient removal.

The effects of different concentrations of graphene oxide (GO) on intracellular metabolism in Chlorella vulgaris (C. vulgaris) and removal of nitrogen and phosphorus nutrients by C. vulgaris from synthetic wastewater were studied. The results demonstrated that cell division of Chlorella vulgaris increased at 24 h and decreased at 96 h after exposure to different concentrations of GO. The removal rates of total nitrogen (TN), ammoniacal nitrogen (NH3-N), phosphate (PO43--P), and chemical oxygen demand (COD) were 24.1%, 70.0%, 37.0%, and 39.6%, respectively, when the concentration of GO was 0.01 mg/L 10 mg/L GO induced severe plasmolysis and cytoplasmic contraction. Furthermore, the protein-like exopolysaccharide (EPS) content of algal cells exposed to 10 mg/L GO decrease to 10.8% of the control group. Simultaneously, the reactive oxygen species (ROS) level was 175.4% of control group. The biological responses to 10 mg/L GO included increase in ROS level, inhibition of saccharide metabolism, and degradation of amino acids. In addition, high concentrations of 10 mg/L GO weakened the carbon fixation process in algal cells. These stress-response behaviors increased cell permeability and oxidative stress. Overall, these findings provide new insights regarding the effects of GO on algal cellular stress responses.

Intensified nitrogen removal in constructed wetlands by novel spray aeration system and different influent COD/N ratios.

The degradation performance of organic matters and microorganism abundance variations of two horizontal subsurface flow constructed wetlands (HSSFCWs) with different influent COD/N ratios and a spray aeration system were investigated. Oxygen-enriched influent water was beneficial to enhance the abilities of nitrification and biodegradation of organic matters. Due to the sufficient carbon source supply under influent COD/N ratio of 8.0, the high removal performances of COD (93%), NH4-N (85%) and total nitrogen (78%) were simultaneously obtained in HSSFCWs with a spray aeration system. The increase in nitrite and nitrate bacteria was beneficial to complete nitrification and improve NH4-N removal. The increase in denitrifying bacteria contributed to complete denitrification and guaranteed the high removal efficiency of NO3-N. The results suggested that the spray aeration combined with high influent COD/N ratios could achieve the high nitrogen removal in HSSFCWs.

Response of submerged macrophytes and periphyton biofilm to water flow in eutrophic environment: Plant structural, physicochemical and microbial properties.

The integrated effects of water flow on submerged macrophytes (Vallisneria natans) and leaf biofilms were comprehensively investigated in eutrophic microcosm. Changes in aquatic environmental factors were analyzed and water flow was found to elevate eutrophic water quality, especially in terms of TP removal. The removal efficiency of TP reached 78.95% in flowing water, which was more than 10-fold higher than in static water. Water flow altered the morphological and physiological characteristics of plants, decreasing the cell wall thickness and rate of photosynthesis, while promoting the accumulation of soluble sugar and protein in leaves. The starch content also increased with water flow, and significantly larger starch granules were observed in chloroplast. Furthermore, oxidative damage was evidenced by the consistently higher content of malondialdehyde in flowing water. Superoxide dismutase (SOD), peroxidase (POD) and Catalase (CAT) were induced in plants exposed to water flow, as an antioxidant stress response. The results of 16S rRNA high-throughput sequencing analysis showed that the structure of the biofilm microbial community changed in response to water flow. These results expand our understanding of the effects of water flow on submerged macrophytes and periphyton biofilms in eutrophic environments.

Effects of carboxyl single-walled carbon nanotubes on synthetic wastewater nutrient removal by an algal-bacterial consortium: Regulation and interaction.

In this study, the morphology, ultrastructure, nutrient removal, metabolite levels, and interaction of an algal-bacterial consortium exposed to different concentrations of carboxylic single-walled carbon nanotubes (C-SWCNT) were investigated. At a C-SWCNT concentration of 0.05 mg·L-1, the removal rates of TN, NH3-N, PO43--P, and COD were 94.7%, 94.8%, 86.4% and 84.3%, respectively. When cells were exposed to 50 mg·L-1 C-SWCNT, its intracellular levels in individual algae and the algal-bacterial consortium were 23.6 µg·g-1 and 12.1 µg·g-1, respectively. C-SWCNT (0.05 mg·L-1) promoted the metabolism of fatty acids, amino acids, small molecules, and acid in the algal-bacterial consortium. The main response to the interaction of C-SWCNT and the consortium was the change in extracellular carbohydrate levels. C-SWCNT also increased chlorophyll a and glycine levels. These findings reveal new insights into our understanding of the biological responses and interactions between C-SWCNT and algal-bacterial consortium.

Nutrient limitation and enzymolysis of phosphorus in Meiliang Bay, Lake Taihu, during algal blooms.

In this study, algal growth potential tests were performed in water samples collected from six sampling sites in Meiliang Bay, Lake Taihu. The potential release of soluble reactive phosphorus (SRP) by enzymatic hydrolysis of enzymatically hydrolyzable phosphorus (EHP) was simultaneously evaluated. Results show that all studied regions were in highly eutrophic states, with additional nitrogen (N) or phosphorus (P) inputs, inducing negligible further increase in algal growth. EHP in water could be rapidly transformed into SRP, further supporting the proliferation of algal blooms. The shortest EHP mineralization time was calculated as 69 minutes; therefore, limiting specific nutrient inputs alone in extremely eutrophic lakes can have a limited effect on suppressing the proliferation of algal blooms. Methods to establish a suitable environmental fate for excessive nitrogen and phosphorus nutrients may be more effective and provide more significant results. PRACTITIONER POINTS: N and P were no longer serving as the limiting factors in Meiliang Bay. Enzymatically hydrolysable phosphorus could be hydrolyzed into soluble reactive phosphorus in a very short period during algal blooms. Both enzymatically hydrolysable phosphorus and soluble reactive phosphorus are required to be curbed in practical eutrophication control.

The concentration thresholds establishment of nitrogen and phosphorus considering the effects of extracellular substrate-to-biomass ratio on cyanobacterial growth kinetics.

Harmful cyanobacterial blooms have caused serious threat to the sustainable development of freshwater ecosystems due to eutrophication, but there is no consensus on nutrients threshold for controlling cyanobacterial bloom. A Monod-based ratio-dependent model was originally developed to investigate the effects of the extracellular substrate-to-biomass (Chlorophyll a) ratio (Sex/X) on the growth kinetics of Cyanobacteria and to determine the concentration thresholds of nitrogen (N) and phosphorus (P) in the form of Sex/X. The results indicated that biomass growth was completely suppressed at Sex/X ≤ 0.21 µg µg-1 for P and Sex/X ≤ 2.82 µg µg-1 for N, which are far lower than the values of most of the eutrophic freshwater lakes. This indicates that in addition to curbing N and P nutrients of eutrophic water, to conduct some biologically mediated changes and to find a suitable environmental fate or a suitable position for excessive N and P nutrients is also of great significance in controlling HABs.

Responses of leaf-associated biofilms on the submerged macrophyte Vallisneria natans during harmful algal blooms.

The present study investigated the physiological responses, photosynthetic activity, and microbial community structure of leaf-associated biofilms on the microphyte Vallisneria natans during a harmful algal bloom. Results of the physiological and photosynthetic indices (Fv/Fm ratios [maximum quantum yield of photosystem II (PSII)]; malondialdehyde content; total chlorophyll; and activities of superoxide dismutase, catalase and peroxidase) indicated that algal blooms could cause inhibition of photosynthesis, oxidative stress and an antioxidant system stress response in Vallisneria natans leaf-associated biofilms. Multifractal analysis suggested that allelochemicals or algal organic matter released by cyanobacteria could reduce the surface roughness of the leaf. Microbial diversity analysis of the biofilms showed that algal blooms slightly altered the microbial community structure while the richness and evenness of the microbial composition remained stable. This study provided useful information to better understand the adverse effects of algal blooms on submerged macrophytes.

Development of model simulation based on BioWin and dynamic analyses on advanced nitrate nitrogen removal in deep bed denitrification filter.

A pilot-scale deep bed denitrification filter using quartz sand as the filter media was operated under filtration velocity of 5.23 m/h. Nitrate, nitrite, ammonia, and total nitrogen removal rates were relatively high at influent C/N ratios of 4:1 and 5:1. A model was developed using software to simulate the processes operating in the filter and improve the related parameters in the actual operations. The normalized sensitivity coefficient and the mean square sensitivity measure were used for the sensitivity analysis. Results showed that the stoichiometric parameters were the most sensitive, which were related to methylotrophs and biofilm. Measured data were consistent with the simulations. Moreover, the order of significance of factors affecting nitrate nitrogen removal was as follows: influent chemical oxygen demand, influent nitrate nitrogen, and hydraulic retention time. Last, the denitrification dynamic model was obtained at influent C/N ratio of 5:1.

The collaborative effect of Chlorella vulgaris-Bacillus licheniformis consortia on the treatment of municipal water.

In this study, the effects of nutrient and dissolved organic matter removal, stress resistance (DNA methylation), and the algae-bacteria dynamic ratio of algal-bacterial consortia in actual municipal wastewater were investigated. Results indicate that the presence of a Chlorella vulgaris-Bacillus licheniformis consortium had profound effects. The removal rates of total nitrogen, ammonium, orthophosphate phosphorus and chemical oxygen demand were 88.82%, 84.98%, 84.87% and 82.25%, respectively. Protein-like substances, which are difficult to degrade in the natural water environment, were significantly degraded in actual municipal wastewater. Furthermore, the microbial diversity was measured. The algal-bacterial consortium did not disrupt the microbial in-situ diversity of the actual municipal wastewater under suitable conditions. The global nuclear DNA methylation level peaked at 7.80%. These results help to understand the effects of algal-bacterial consortia on nutrient and pollutant removal and adaptability in actual municipal wastewater.

The interactions of algae-bacteria symbiotic system and its effects on nutrients removal from synthetic wastewater.

The ability of Chlorella vulgaris-Bacillus licheniformis and Microcystis aeruginosa-Bacillus licheniformis consortiums to eliminate total dissolved nitrogen (TDN), total dissolved phosphorus (TDP), and soluble chemical oxygen demand (sCOD) from synthetic wastewater was studied. The highest values of dry cell weight, chlorophyll-a, and chlorophyll metabolism related genes/bacterial rRNA gene copies were obtained in the Chlorella vulgaris-Bacillus licheniformis system at Chlorella vulgaris and Bacillus licheniformis ratio of 1:3. On the 10th day, the Chlorella vulgaris-Bacillus licheniformis system at this ratio removed 86.55%, 80.28% and 88.95% of sCOD, TDP and TDN, respectively. But, the Microcystis aeruginosa-Bacillus licheniformis system at this ratio only removed 65.62%, 70.82%, and 21.56% of sCOD, TDP and TDN, respectively. Chlorella vulgaris and Bacillus licheniformis could coexist as an algae-bacteria consortia and quorum sensing substances (autoinducing peptides and bis (3'-5') diguanylic acid) concentrations were measured. Finally, the interactions and communication patterns between Chlorella vulgaris and Bacillus licheniformis were depicted.