Genetic response analysis of Beauveria bassiana Z1 under high concentration Cd(II) stress.

Cadmium (Cd(II)) has carcinogenic and teratogenic toxicity, which can be accumulated in the human body through the food chain, endangering human health and life. In this study, a highly Cd(II)-tolerant fungus named Beauveria bassiana Z1 was studied, and its Cd(Ⅱ) removal efficiency was 71.2% when the Cd(II) concentration was 10 mM. Through bioanalysis and experimental verification of the transcriptome data, it was found that cadmium entered the cells through calcium ion channels, and then complexed with intracellular glutathione (GSH) and stored in vacuoles or excluded extracellular by ABC transporters. Cytochrome P450 was significantly upregulated in many pathways and actively participated in detoxification related reactions. The addition of cytochrome inhibitor taxifolin reduced the removal efficiency of Cd(II) by 45%. In the analysis, it demonstrated that ACOX1 gene and OPR gene of jasmonic acid (JA) synthesis pathway were significantly up-regulated, and were correlated with bZIP family transcription factors cpc-1_0 and pa p1_0. The results showed that exogenous JA could improve the removal efficiency of Cd(II) by strain Z1.

Metabolic diversity shapes vegetation-enhanced methane oxidation in landfill covers: Multi-omics study of rhizosphere microorganisms.

Vegetation root exudates have the ability to shape soil microbial community structures, thereby enhancing CH4 bio-oxidation capacity in landfill cover systems. In this study, the CH4 oxidation capacity of indigenous vegetation rhizosphere microorganisms within operational landfill covers in Chongqing, China, was investigated for the first time, with the objective of identifying suitable plant candidates for CH4 mitigation initiatives within landfill cover systems. Furthermore, a multi-omics methodology was employed to explore microbial community structures and metabolic variances within the rhizospheric environment of diverse vegetation types. The primary aim was to elucidate the fundamental factors contributing to divergent CH4 oxidation capacities observed in rhizosphere soils. The findings demonstrated that herbaceous vegetation predominated in landfill covers. Notably, Rumex acetosa exhibited the highest CH4 oxidation capacity in the rhizosphere soil, approximately 20 times greater than that in non-rhizosphere soil. Root exudates played a crucial role in inducing the colonization of CH4-oxidizing functional microorganisms in the rhizosphere, subsequently prompting the development of specific metabolic pathways. This process, in turn, enhanced the functional activity of the microorganisms while concurrently bolstering their tolerance to microbial pollutants. Consequently, the addition of substances like Limonexic acid strengthened the CH4 bio-oxidation process, thereby underscoring the suitability of Rumex acetosa and similar vegetation species as preferred choices for landfill cover vegetation restoration.

Quantitative analysis of TCE biodegradation pathway in landfill cover utilizing continuous monitoring, droplet digital PCR and multi-omics sequencing technology.

The remediation of volatile chlorinated hydrocarbons in the quasi-vadose zone has become a significant challenge. We applied an integrated approach to assess the biodegradability of trichloroethylene to identify the biotransformation mechanism. The formation of the functional zone biochemical layer was assessed by analyzing the distribution of landfill gas, physical and chemical properties of cover soil, spatial-temporal variations of micro-ecology, biodegradability of landfill cover soil and distributional difference metabolic pathway. Real-time online monitoring showed that trichloroethylene continuously undergoes anaerobic dichlorination and simultaneous aerobic/anaerobic conversion-aerobic co-metabolic degradation on the vertical gradient of the landfill cover system and reduction in trans-1,2-dichloroethylene in the anoxic zone but not 1,1-dichloroethylene. PCR and diversity sequencing revealed the abundance and spatial distribution of known dichlorination-related genes within the landfill cover, with 6.61 ± 0.25 × 104-6.78 ± 0.09 × 106 and 1.17 ± 0.78 × 103-7.82 ± 0.07 × 105 copies per g/soil of pmoA and tceA, respectively. In addition, dominant bacteria and diversity were significantly linked with physicochemical factors, and Mesorhizobium, Pseudoxanthomonas and Gemmatimonas were responsible for biodegradation in the aerobic, anoxic and anaerobic zones. Metagenome sequencing identified 6 degradation pathways of trichloroethylene that may occur in the landfill cover; the main pathway was incomplete dechlorination accompanied by cometabolic degradation. These results indicate that the anoxic zone is important for trichloroethylene degradation.

Microecological health assessment of water environment and sediment based on metagenomics: a case study of Guixi River in Chongqing, China.

River sediment is vital in containing water pollution and strengthening water remediation. This paper has conducted a study on the microecological health assessment of the sediment and water body of Guixi River in Dianjiang, Chongqing, China, using metagenomics sequencing and microbial biological integrity index (M-IBI) technology. The analysis of physical and chemical characteristics shows that the concentration of TN varies from 2.62 to 9.76 mg/L in each sampling section, and the eutrophication of the water body is relatively severe. The proportion of Cyanobacteria in the sampling section at the sink entrance is higher than that of other sites, where there are outbreaks of water blooms and potential hazards to human health. The dominant functions of each site include carbon metabolism, TCA cycle, and pyruvate metabolism. In addition, the main virulence factors and antibiotic resistance genes in sediment are Type IV pili (VF0082), LOS (CVF494), MymA operon (CVF649), and macrolide resistance genes macB, tetracyclic tetA (58), and novA. Correlation analysis of environmental factors and microorganisms was also performed, and it was discovered that Thiothrix and Acidovorax had obvious gene expression in the nitrogen metabolism pathway, and the Guixi River Basin had a self-purification capacity. Finally, based on the microecological composition of sediment and physical and chemical characteristics of the water body, the health assessment was carried out, indicating that the main pollution area was Dianjiang Middle School and the watershed near the sewage treatment plant. The findings should theoretically support an in-depth assessment of the water environment's microecological health.

Domestication and pilot-scale culture of mixed bacteria HY-1 capable of heterotrophic nitrification-aerobic denitrification.

To further investigate the potential of heterotrophic nitrification-aerobic denitrification (HN-AD) bacteria for practical applications, the HN-AD mixed bacteria HY-1 were enriched and domesticated in this study. After five generations of domestication, the mixture was able to remove 98% of ammonia nitrogen (400 mg/L) and 81.9% of mixed nitrogen source (nitrate, nitrite). Changes in community structure in the domestication process of mixed microorganisms were studied using 16S rDNA-seq. The results indicated an increase in the abundance of Acinetobacter from 16.9% to 80%. The conditions for the expanded culture of the HY-1 were also optimized. Moreover, A pilot-scale expanded reactor with a capacity of 1000L was constructed, and the HY-1 was successfully expanded from 0.1L to 800L. The community structures of the HY-1 remained stable after the expanded culture, with Acinetobacter as the dominant species. Moreover, the HY-1 demonstrated adaptability to actual high ammonia nitrogen wastewater and showed potential for practical application.

Water microecology is affected by seasons but not sediments: A spatiotemporal dynamics survey of bacterial community composition in Lake Changshou-The largest artificial lake in southwest China.

This study aimed to evaluate the correlation between microecology of sediments and water as well as their spatial-temporal variations in Changshou Lake. The results demonstrated that microecology in the lake exhibits spatiotemporal heterogeneity, and microbial diversity of sediments was significantly higher than that of water body. Further, it was found that there was statistically insignificant positive correlation between microecology of sediments and that of water body. PCoA and community structure analysis revealed that the predominant phyla which exhibited significant spatial differences in sediments were Proteobacteria, Actinobacteria and Planctomycetes. While, the distribution of dominant bacteria Actinobacteria and Verrucomicrobia in water body showed significant seasonal differences. Microbial networks analysis indicated that there was a cooperative symbiotic relationship between lake microbial communities. Notably, the same bacterial genus had no significant positive correlation in sediment and water, which suggested that bacteria transport between sediment-water interface does not influence the microecological functions of lake water.

Effect mechanism of individual and combined salinity on the nitrogen removal yield of heterotrophic nitrification-aerobic denitrification bacteria.

One of the biggest challenges of applying heterotrophic nitrification-aerobic denitrification (HN-AD) bacteria to treat high salt organic wastewater lies in the inhibitory effect exerted by salinity. To study the inhibition effect and underlying mechanism induced by different ion types and ion composition, the individual and combined effects of NaCl, KCl and Na2SO4 on HN-AD bacteria Acinetobacter sp. TAC-1 were systematically investigated by batch experiments. Results indicated that the ammonia nitrogen removal yield and TAC-1 activity decreased with increased salt concentration. NaCl, KCl and Na2SO4 exerted different degrees of inhibition on TAC-1, with half concentration inhibition constant values of 0.205, 0.238 and 0.110 M, respectively. A synergistic effect on TAC-1 was found with the combinations of NaCl + KCl, NaCl + Na2SO4 and NaCl + KCl + Na2SO4. The whole RNA resequencing suggested that transcripts of denitrification genes (nirB and nasA) were significantly downregulated with increased Na2SO4 concentration. Simultaneously, Na2SO4 stress disrupted cell respiration, DNA replication, transcription, translation, and induced oxidative stress. Finally, we proposed a conceptual model to summarize the inhibition mechanisms and possible response strategies of TAC-1 bacteria under Na2SO4 stress.

Nitrogen removal and mechanism of an extremely high-ammonia tolerant heterotrophic nitrification-aerobic denitrification bacterium Alcaligenes faecalis TF-1.

A novel heterotrophic nitrifying bacterium with high salt and high ammonia nitrogen tolerance, Alcaligenes faecalis TF-1, was isolated from the leachate of a landfill. The verification of nitrogen removal efficiency of different nitrogen sources and PCR amplification electrophoresis results showed that the HN-AD pathway of the strain TF-1 was NH4+ â†’ NH2OH â†’ NO â†’ N2O â†’ N2. The results of parameter optimization showed that the optimal nitrogen removal conditions were as follows: sodium citrate as carbon source, C/N = 16, pH = 7, and NH4+-N loading of 808.21 mg/L. The strain TF-1 could remove about 94.60% of ammonia nitrogen (1963.94 mg/L). The salinity tolerance range of the strain TF-1 was 0-70 g/L, and the removal efficiency was 52.87% at salinity 70 g/L and NH4+-N concentration 919.20 mg/L and 55.67% at pH = 10 and NH4+-N concentration 994.82 mg/L. The extreme environmental adaptability and remarkable nitrogen removal performance make this strain a promising candidate in leachate treatment.

Characterization of a novel salt-tolerant strain Sphingopyxis sp. CY-10 capable of heterotrophic nitrification and aerobic denitrification.

A novel heterotrophic nitrification and aerobic denitrification (HN-AD) strain CY-10 was isolated and identified as Sphingopyxis sp. When ammonium, nitrate or nitrite was used as the sole nitrogen source (300 mg/L), the maximum nitrogen removal efficiency of strain CY-10 were 100%, 91.1% and 68.5%, respectively. The optimal salinity for ammonia nitrogen removal by strain CY-10 was in the range of 0-5%. At the salinity of 5%, a maximum nitrogen removal rate of 6.25 mg/(L·h) was realized. Metabonomics data showed that the metabolic levels of sucrose and D-tagatose increased significantly at 5% salinity condition, enabling the strain to regulate osmotic pressure and survive in high-salt environments. Functional genes were successfully amplified by quantitative PCR, and HN-AD pathway of strain CY-10 followed NH4+-N â†’ NH2OH â†’ NO2--N â†’ NO â†’ N2O â†’ N2. These findings show that strain CY-10 has great potential in nitrogen removal treatment of saline wastewater.

[Transformation mechanism of carbon tetrachloride and the associated micro-ecology in landfill cover, a typical functional layer zone].

Landfill is one of the important sources of carbon tetrachloride (CT) pollution, and it is important to understand the degradation mechanism of CT in landfill cover for better control. In this study, a simulated landfill cover system was set up, and the biotransformation mechanism of CT and the associated micro-ecology were investigated. The results showed that three stable functional zones along the depth, i.e., aerobic zone (0-15 cm), anoxic zone (15-45 cm) and anaerobic zone (> 45 cm), were generated because of long-term biological oxidation in landfill cover. There were significant differences in redox condition and microbial community structure in each zone, which provided microbial resources and favorable conditions for CT degradation. The results of biodegradation indicated that dechlorination of CT produced chloroform (CF), dichloromethane (DCM) and Cl- in anaerobic and anoxic zones. The highest concentration of dechlorination products occurred at 30 cm, which were degraded rapidly in aerobic zone. In addition, CT degradation rate was 13.2-103.6 µg/(m2·d), which decreased with the increase of landfill gas flux. The analysis of diversity sequencing revealed that Mesorhizobium, Thiobacillus and Intrasporangium were potential CT-degraders in aerobic, anaerobic and anoxic zone, respectively. Moreover, six species of dechlorination bacteria and eighteen species of methanotrophs were also responsible for anaerobic transformation of CT and aerobic degradation of CF and DCM, respectively. Interestingly, anaerobic dechlorination and aerobic transformation occurred simultaneously in the anoxic zone in landfill cover. Furthermore, analysis of degradation mechanism suggested that generation of stable anaerobic-anoxic-aerobic zone by regulation was very important for the harmless removal of full halogenated hydrocarbon in vadose zone, and the increase of anoxic zone scale enhanced their removal. These results provide theoretical guidance for the removal of chlorinated pollutants in landfills.

Direct aerobic oxidation (DAO) of chlorinated aliphatic hydrocarbons: A review of key DAO bacteria, biometabolic pathways and in-situ bioremediation potential.

Contamination of aquifers and vadose zones with chlorinated aliphatic hydrocarbons (CAH) is a world-wide issue. Unlike other reactions, direct aerobic oxidation (DAO) of CAHs does not require growth substrates and avoids the generation of toxic by-products. Here, we critically review the current understanding of chlorinated aliphatic hydrocarbons-DAO and its application in bioreactors and at the field scale. According to reports on chlorinated aliphatic hydrocarbons-DAO bacteria, isolates mainly consisted of Methylobacterium and Proteobacterium. Chlorinated aliphatic hydrocarbons-DAO bacteria are characterized by tolerance to a high concentration of CAHs and highly efficient removal of CAHs. Trans-1,2-dichloroethylene (t-DCE) is easily transformed biomass for bacteria, followed by 1,2-dichloroethane (1,2-DCA), dichloromethane (DCM), vinyl chloride (VC) and cis-1,2-dichloroethylene (c-DCE). Significant differences in the maximum specific growth rates were observed with different CAHs and biometabolic pathways for DCM, 1,2-DCA, VC and c-DCE degradation have been successfully parsed. Detection of the functional genes etnC and etnE is useful for the determination of active VC DAO bacteria. Additionally, DAO bacteria have been successfully applied to CAHs in new types of bioreactors with satisfactory results. To the best of the authors' knowledge, only one study on DAO-CAHs was conducted in-situ and resulted in 99% CAH removal. Lastly, we put forward future development prospect of chlorinated aliphatic hydrocarbons-DAO.

Response of rotating biological contactor started up by heterotrophic nitrification-aerobic denitrification bacteria to various C/N ratios.

To improve the low pollutant removal efficiency of traditional biological methods for treating livestock and poultry breeding wastewater under a relatively low temperature, a rotating biological contactor (RBC) inoculated with heterotrophic nitrification-aerobic denitrification (HN-AD) bacteria was designed. A quick start-up process and a well removal performance had been achieved in the novel RBC. To elucidate the anti-load shock ability of the novel RBC, the effects of C/N ratio on nitrogen removal and microbial assemblage were focused under a low temperature (12 ± 2 °C). Results showed that the highest NH4+-N and TN removal efficiency were 99.57 ± 0.31% and 68.41 ± 0.52%. Microbial diversity analysis based on high throughput sequencing technique showed that Arcobacter and Flavobacterium with an increasing relative abundance were the key to ensure high nitrogen removal efficiently at a low C/N ratio and temperature. Moreover, nitrogen transferring pathways of the novel RBC was revealed and dissimilatory nitrate reduction and denitrification were the main pathways. The excellent pollutant removal performance demonstrates that the novel RBC is a promising process to effectively treat wastewater with low C/N ratio and low temperature.

Cometabolic degradation mechanism and microbial network response of methanotrophic consortia to chlorinated hydrocarbon solvents.

The cometabolism mechanism of chlorinated hydrocarbon solvents (CHSs) in mixed consortia remains largely unknown. CHS biodegradation characteristics and microbial networks in methanotrophic consortia were studied for the first time. The results showed that all CHSs can efficiently be degraded via cometabolism with a maximum degradation rate of 4.8 mg/(h·gcell). Chloroalkane and chloroethylene were more easily degraded than chlorobenzenes by methanotrophic consortia, especially nonfully chlorinated aliphatic hydrocarbons, which were converted to Cl- with a production rate of 0.29-0.36 mg/(h·gcell). In addition, the microecological response results indicated that Methylocystaceae (49.0%), Methylomonas (65.3%) and Methylosarcina (41.9%) may be the major functional degraders in methanotrophic consortia. Furthermore, the results of the microbial correlation network suggested that interactive relationships constructed by type I methanotrophs and heterotrophs determined biodegradability. Additionally, PICRUSt analysis showed that CHSs could increase the relative abundance of CHS degradation genes and reduce the relative abundance of methane oxidation genes, which was in good agreement with the experimental results.

Effect of bacteria-to-algae volume ratio on treatment performance and microbial community of a novel heterotrophic nitrification-aerobic denitrification bacteria-chlorella symbiotic system.

A novel symbiotic system combined by heterotrophic nitrification-aerobic denitrification (HN-AD) mixed bacteria and Chlorella pyrenoidosa was firstly proposed to resolve the poor tolerance and nitrogen removal performance of traditional symbiotic system for treating high ammonia biogas slurry. Results showed that the volume ratio of bacteria to algae had significant effects on nitrogen removal efficiency, microbial community structure, functional bacteria and genes. The optimal ratio was 1/3, and the average removal efficiency of TN and TP increased by 28.9% and 67.6% respectively, compared to those of HN-AD bacteria. High-throughput sequencing indicated nitrogen removal was jointly completed by HN-AD and heterotrophic denitrification. HN-AD bacteria Halomonas and Pseudomonas played a key role in nitrogen removal, and Rhodocyclaceae and Paracoccus took an important part in phosphorus removal. According to the functional gene prediction, the total relative abundance of nitrogen removal genes (0.0127%) and narG, narH and narL genes (0.0054%) were highest in 1/3 system.

Efficiency and microbial diversity of aeration solid-phase denitrification process bioaugmented with HN-AD bacteria for the treatment of low C/N wastewater.

To evaluate the simultaneous nitrification and denitrification (SND) performance of the aeration solid-phase denitrification (SPD) process and improve the operating efficiency, aeration SPD process using polybutanediol succinate as carbon source was optimized and the process was bioaugmented with heterotrophic nitrification-aerobic denitrification bacteria for the treatment of real wastewater. The results showed that after bioaugmentation, the total nitrogen removal efficiency of the aeration SPD process increased by 50.46 % under condition of dissolved oxygen (DO) 3 mg/L. According to Illumina MiSeq sequencing and correlation analyses, the microbial community can perform SND under the conditions of DO 5 mg and HRT 6 h, but is susceptible to DO. Bioaugmentation mainly affected the carbon source metabolic network with heterotrophic bacteria Methyloversatilis, Thiothrix, and norank_Lentimicrobiaceae as nodes to change the community structure, thereby improving the performance of the functional microbial community. Kyoto Encyclopedia of Genes and Genomes analysis suggested that narB, narG, narH, nirK and narI were the key genes involved in the response to bioaugmentation. This work provides new insights for the application of the SPD process in wastewater treatment.

Heterotrophic nitrification and aerobic denitrification by a novel Acinetobacter sp. TAC-1 at low temperature and high ammonia nitrogen.

A novel strain was isolated from swinewastewater and identified as Acinetobacter sp. TAC-1 based on its phylogenetic and phenotypic characteristics. The strain TAC-1 was found to have a high ability to metabolize ammonium-N under low temperature condition. The strain TAC-1 could remove approximately 94.6% of ammonium-N (400 mg/L), 93.3% of nitrate-N (400 mg/L) and 42.4% of nitrite-N (400 mg/L) at 5 °C. The functional genes nitrate reductase gene (narG) and nitrite reductase gene (nirK, nirS) were successfully amplified by qPCR, further evidencing the heterotrophic nitrification and aerobic denitrification capability of Acinetobacter sp. TAC-1. The transcriptome data confirmed that the membrane transport protein and unsaturated fatty acid dehydrogenase-related genes of the strain TAC-1 were significantly up-regulated at 5 °C, enabling it to survive low temperatures. The high nitrogen removal ability at 5 °C makes this strain have a good application prospect.

Response of wastewater treatment performance, microbial composition and functional genes to different C/N ratios and carrier types in MBBR inoculated with heterotrophic nitrification-aerobic denitrification bacteria.

To operate the moving bed biofilm reactor inoculated with HN-AD bacteria (B-MBBR) instead of activated sludge for livestock and poultry breeding wastewater (LPBW) disposal in most efficient manner, nitrogen removal (NR) efficiency and microbial composition of two MBBRs with different carrier types under various C/N ratios were explored. Results indicated that the performance on NR greatly various in different carrier types under various C/N ratios. Attributing to the bacterial protection provided by the porous structure of polyvinyl alcohol (PVA) gel, MBBR using PVA gel as the carrier exhibited a more stable NR performance (range from 78.05% to 83.76%) versus that using Kaldnes (K1) as the carrier (range from 78.05% to 83.76%). Besides, microbial analysis indicated that MBBR with PVA gel as the carrier is conducive to the growth of oligotrophic and HN-AD bacteria (Paracoccus and Acinetobacter), and the highest relative abundance was 16.37% at C/N ratio of 6.

Progress of carbon-based electrocatalysts for flexible zinc-air batteries in the past 5 years: recent strategies for design, synthesis and performance optimization.

The increasing popularity of wearable electronic devices has led to the rapid development of flexible energy conversion systems. Flexible rechargeable zinc-air batteries (ZABs) with high theoretical energy densities demonstrate significant potential as next-generation flexible energy devices that can be applied in wearable electronic products. The design of highly efficient and air-stable cathodes that can electrochemically catalyze both the oxygen reduction reaction (ORR) and oxygen evolution reaction (OER) are highly desirable but challenging. Flexible carbon-based catalysts for ORR/OER catalysis can be broadly categorized into two types: (i) self-supporting catalysts based on the in situ modification of flexible substrates; (ii) non-self-supporting catalysts based on surface coatings of flexible substrates. Methods used to optimize the catalytic performance include doping with atoms and regulation of the electronic structure and coordination environment. This review summarizes the most recently proposed strategies for the synthesis of designer carbon-based electrocatalysts and the optimization of their electrocatalytic performances in air electrodes. And we significantly focus on the analysis of the inherent active sites and their electrocatalytic mechanisms when applied as flexible ZABs catalysts. The findings of this review can assist in the design of more valuable carbon-based air electrodes and their corresponding flexible ZABs for application in wearable electronic devices.

Biodegradation of phenol by a highly tolerant strain Rhodococcus ruber C1: Biochemical characterization and comparative genome analysis.

A novel phenol-degrading strain was isolated and identified as Rhodococcus ruber C1. The degradation analysis shows that 1806 mg/L of phenol can be completely degraded by strain C1 within 38 h, and the maximum specific growth rate (µmax=1.527 h-1) and maximum specific phenol degradation rate (qmax=3.674 h-1) indicate its excellent phenol metabolism capability. More importantly, phenol can be degraded by strain C1 in the temperature range of 20-45 °C within 72 h, and with longer degradation time, phenol can be completely degraded even at 10, 15 and 50 °C. The whole genome of strain C1 was sequenced, and a comparative genome analysis of strain C1 with 36 other genomes of Rhodococcus was performed. A remarkable gene family expansion occurred during the evolution of Rhodococcus, and a comprehensive evolutionary picture of Rhodococcus at genomic level was presented. Moreover, the copy number of genes involved in phenol metabolism was compared among genus Rhodococcus, and the results demonstrate high phenol degradation capability of strain C1 at genomic level. These findings suggest that Rhodococcus ruber C1 is a bacterium capable of degrading phenol efficiently in the temperature range of 10-50 °C.

Removal of CX3R-type disinfection by-product precursors from rainwater with conventional drinking water treatment processes.

In addition to surface water and groundwater, rainwater is used as an important drinking water source in many parts of the world, especially in areas with serious water pollution or insufficient water resources. Conventional drinking water treatment technologies can remove dissolved organic matter and therefore reduce the formation of disinfection by-products (DBPs) during subsequent disinfection using surface water or groundwater as drinking water sources. However, little information has been known about the effect of conventional water treatment processes on DBP formation when rainwater is used as drinking water source. This study evaluated CX3R-type DBP precursors removal from rainwater by conventional drinking water treatments and the corresponding decrease of CX3R-type DBP (trihalomethanes (THMs), haloaldehydes (HALs), haloacetonitriles (HANs) and haloacetamides (HAMs)) formation and toxicity during the subsequent chlor(am)ination. The result showed that both sand filtration (SF) and activated carbon filtration (GAC) were able to remove DBP precursors and GAC outperformed SF, but no DBP precursors removal was observed during coagulation-sedimentation treatment. Among all treatments, SF + GAC was the most effective for DBP precursors removal, with removal efficiencies of 64.2% DOC, 98% DON and 76.6% UV254. Correspondingly, both SF and GAC decreased the formation of THMs, HALs, HANs and HAMs, and GAC performed better than SF. The combination of SF and GAC, especially SF + GAC, greatly decreased DBP formation, with average reduction of 79.2% and 85% during chlorination and chloramination respectively. After different treatments, the comprehensive toxicity risk of CX3R-type DBPs was all reduced, among which GAC + SF exhibited superior performance. Generally, the main contribution of integrated toxicity was HANs during chlor(am)ination. The formation potential of THMs, HALs, HANs and HAMs and the corresponding integrated toxicity were greater during chlorination than that during chloramination. Therefore, the combination of GAC and chloramination was promising in mitigating the comprehensive toxicity risk of THMs, HALs, HANs and HAMs for rainwater.