Metagenomic insights into the profile of antibiotic resistomes in sediments of aquaculture wastewater treatment system

To meet the rapidly growing global demand for aquaculture products, large amounts of antibiotics were used in aquaculture, which might accelerate the evolution of antibioticresistant bacteria(ARB) and the propagation of antibiotic genes(ARGs). In our research, we revealed the ARGs profiles, their co-occurrence with mobile genetic elements(MGEs), and potential hosts in sediments of a crab pond wastewater purification system based on metagenomic analysis. The residual antibiotic seems to increase ...     

Metabolic responses of indigenous bacteria in chicken faeces and maggots to multiple antibiotics via heavy water labeled single-cell Raman spectroscopy

The use of maggots derived from chicken faeces as fish diets might serve as a vehicle for the widespread of multiple antibiotic resistant bacteria(ARB) in the environment. Heavy water labeled single-cell Raman spectroscopy(D 2 O-Raman) was applied to detect the metabolic responses of indigenous bacteria in chicken faeces and maggots to different concentrations of combined colistin, kanamycin, and vancomycin. By incubating the samples with D 2 O and antibiotics, metabolically active bacterial cel...     

Comparison of different disinfection processes in the effective removal of antibiotic-resistant bacteria and genes

This study compared three different disinfection processes(chlorination, E-beam, and ozone) and the efficacy of three oxidants(H2O2, S2O-8, and peroxymonosulfate(MPS)) in removing antibiotic resistant bacteria(ARB) and antibiotic resistance genes(ARGs) in a synthetic wastewater. More than30 mg/L of chlorine was needed to remove over 90% of ARB and ARG. For the E-beam method, only1 dose(kGy) was needed to remove ARB and ARG, and ozone could reduce ARB and ARG by more than 90% even at 3 mg/L ozone concentration. In the ozone process, CT values(concentration × time)were compared for ozone alone and combined with different catalysts based on the 2-log removal of ARB and ARG. Ozone treatment yielded a value of 31 and 33(mg·min)/L for ARB and ARGs respectively. On the other hand, ozone with persulfate yielded 15.9 and 18.5(mg·min)/L while ozone with monopersulfate yielded a value of 12 and 14.5(mg·min)/L. This implies that the addition of these catalysts significantly reduces the contact time to achieve a 2-log removal, thus enhancing the process in terms of its kinetics.     

Isolation and characterization of a thermophilic Bacillus shackletonii K5 from a biotrickling filter for the production of polyhydroxybutyrate

Polyhydroxyalkanoates（PHAs） are aliphatic polyesters accumulated intracellularly by both Gram-negative and Gram-positive bacteria. However, compared to the PHAs of Gramnegative bacteria, few endotoxins（lipopolysaccharides, LPS）, which would be co-purified with PHAs and cause immunogenic reactions, are found in the PHAs produced by Gram-positive bacteria. A thermophilic Gram-positive bacterium K5, which exhibited good growth and polyhydroxybutyrate（PHB）-accumulating ability, has been isolated and characterized from a biotrickling filter designed for the removal of NOx from flue gas in a coal-fired power plant in China. Based on the biochemical characterization and 16 S rRNA gene sequence（Genbank accession no. JX437933）, the strain K5 has been identified as Bacillus shackletonii, which has rarely been reported in the literature, and this report is the first time that B. shackletonii has been found to accumulate PHB. The strain K5 was able to utilize glucose as carbon source to synthesize PHB at a broad range of temperatures（from 35 to 50 °C）, and the ideal temperature was 45 °C. The strain K5 could effectively yield PHB of up to 69.9% of its cell dry weight（CDW）（2.28 g/L） in flask experiments employing glucose as carbon source at 45 °C, followed by 56.8% and 52.3% of its CDW when using sodium succinate and glycerol as carbon source, respectively. For batch cultivation, the strain K5 was able to produce PHB of up to 72.6% of its cell dry weight（9.76 g/L） employing glucose as carbon source at 45 °C and pH 7.0.     

Microbial community variation in phytoremediation of triazophos by Canna indica Linn. in a hydroponic system

Phytoremediation of triazophos (O,O-diethyl-O-(1-phenyl-1,2,4-triazole-3-base) sulfur phosphate, TAP) pollution by Canna indica Linn. in a hydroponic system has been well studied, whereas the microbial mechanism on TAP degradation is still unknown. The variation in microbial community compositions was investigated by analyzing phospholipid fatty acids (PLFAs) profiles in microbes under TAP exposure. The TAP exposure resulted in an increase in proportions of fatty acid 16:0 and decrease in fatty acid 18:2ω9,12c, indicating that TAP may stimulate the reproduction of microorganisms and inhibit the growth of fungi to some degree. Significant correlation was found between the ratio of fungi to bacteria and TAP removal (r 2 = 0.840, p 0.01). In addition, the microbial community in the phytoremediation system with C. indica was dominated by Gram negative bacteria, which possibly contributed to the degradation of TAP. These results indicated that TAP might induce the colonization of bacteria in the hydroponic system planted with C. indica, and lead to a discrimination of microbial community, which might be one of the mechanisms on TAP dissipation in phytoremediation system.     

Difference in calcium ion precipitation between free and immobilized Halovibrio mesolongii HMY2

Biomineralization has become a research focus in wastewater treatment due to its much lower costs compared to traditional methods. However, the low sodium chloride (NaCl)-tolerance of bacteria limits applications to only water with low NaCl concentrations. Here, calcium ions in hypersaline wastewater (10% NaCl) were precipitated by free and immobilized Halovibrio mesolongii HMY2 bacteria and the differences between them were determined. The results show that calcium ions can be transformed into several types of calcium carbonate with a range of morphologies, abundant organic functional groups (C-H, C-O-C, C=O, etc), protein secondary structures (β-sheet, α-helix, 3₁₀ helix, and β-turn), P=O and S-H indicated by P2p and S2p, and more negative δ¹³C_PDB (‰) values (-16.8‰ to -18.4‰). The optimal conditions for the immobilized bacteria were determined by doing experiments with six factors and five levels and using response surface method. Under the action of two groups of immobilized bacteria prepared under the optimal conditions, by the 10^th day, Ca²⁺ ion precipitation ratios had increased to 79%-89% and 80%-88% with changes in magnesium ion cencentrations. Magnesium ions can significantly inhibit the calcium ion precipitation, and this inhibitory effect can be decreased under the action of immobilized bacteria. Minerals induced by immobilized bacteria always aggregated together, had higher contents of Mg, P, and S, lower stable carbon isotope values and less well-developed protein secondary structures. This study demonstrates an economic and eco-friendly method for recycling calcium ions in hypersaline wastewater, providing an easy step in the process of desalination.     

Influence of metal ions on sulfonamide antibiotics biochemical behavior in fiber coexisting system

Metal ions and fiber are common compounds in the livestock and poultry manure,which will affect the fate of organic compounds in aqueous environment. However,limited research has addressed the effect of coexisting metal ions and fiber on the biodegradation of sulfonamide antibiotics. Accordingly, a compositing study was performed to assess the effect of metal ions(Fe3+and Cu2+) on the biodegradation of sulfadimethoxine sodium salt(SDM) in the presence of fiber. The enhanced adsorption of SDM onto fiber in the presence of metal ions can be attributed to the π+–π electron donor acceptor(EDA) interaction. The microbial(Phanerochaete chrysosprium) could easily attach onto fiber forming attached microbial, and the degradation rates of SDM of immobilized bacteria in the presence of Fe3 +were 100%, which were significantly higher than those of free bacteria(45%). This study indicates that Fe3 +and fiber could enhance the biodegradation of SDM. Fiber acts as adsorbent, carrier, and substrate which enhanced the removal of SDM.     

Stable-isotope probing of bacterial community for dissolved inorganic carbon utilization in Microcystis aeruginosa-dominated eutrophic water

Dissolved inorganic carbon (DIC) is an important source of carbon in aquatic ecosystems, especially under conditions of increased frequency of cyanobacterial bloom. However, the importance of bacteria in direct or indirect utilization of DIC has been widely overlooked in eutrophic freshwater. To identify the functional bacteria that can actively utilize DIC in eutrophic freshwater during cyanobacterial bloom, stable-isotope probing (SIP) experiments were conducted on eutrophic river water with or without inoculation with cyanobacteria (Microcystis aeruginosa). Our 16S rRNA sequencing results revealed the significance of Betaproteobacteria, with similar relative abundance as Alphaproteobacteria, in the active assimilation of H¹³CO₃^? into their DNA directly or indirectly, which include autotrophic genera Betaproteobacterial ammonia-oxidizing bacteria. Other bacterial groups containing autotrophic members, e.g. Planctomycetes and Nitrospira, also presented higher abundance among free-living bacteria in water without cyanobacteria. Microcystis aggregates showed a preference for some specific bacterial members that may utilize H¹³CO₃^? metabolized by Microcystis as organic matter, e.g. Bacteroidetes (Cytophagales, Sphingobacteriales), and microcystin-degrading bacteria Betaproteobacteria (Paucibacter/Burkholderiaceae). This study provides some valuable information regarding the functional bacteria that can actively utilize DIC in eutrophic freshwater.     

Cd2+ removal from wastewater by sulfate reducing bacteria with an anaerobic fluidized bed reactor

A process of treatment for containing Cd²⁺ wastewater by sulfate reducing bacteria with upflow anaerobic fluidized bed reactor has been studied. When the concentration of COD and Cd²⁺ in the influent were 270 5mg/L and 100mg/L respectively and hydraulic retention time was 4 hours, the removal rate of COD and Cd²⁺ were higher than 73 8% and 99 8% respectively. The reactor can treat as high as 1000mg/L of concentration of Cd²⁺ . The highest removal velocity rate of Cd²⁺ reached 2999 1mg/(L·d). And the possible relationship between sulfate reducing bacteria and methanogenic bacteria was discussed.     

Coupled effects of methane monooxygenase and nitrogen source on growth and poly-β-hydroxybutyrate (PHB) production of Methylosinus trichosporium OB3b

The coupled effects of nitrogen source and methane monooxygenase(MMO) on the growth and poly-β-hydroxybutyrate(PHB) accumulation capacity of methanotrophs were explored.The ammonia-supplied methanotrophs expressing soluble MMO(s MMO) grew at the highest rate, while N_2-fixing bacteria expressing particulate MMO(p MMO) grew at the lowest rate. Further study showed that more hydroxylamine and nitrite was formed by ammonia-supplied bacteria containing p MMO, which might cause their slightly lower growth rate. The highest PHB content(51.0%) was obtained under nitrogen-limiting conditions with the inoculation of nitrate-supplied bacteria containing p MMO. Ammoniasupplied bacteria also accumulated a higher content of PHB(45.2%) with the expression of p MMO, while N_2-fixing bacteria containing p MMO only showed low PHB production capacity(32.1%). The maximal PHB contents of bacteria expressing s MMO were low, with no significant change under different nitrogen source conditions. The low MMO activity,low cell growth rate and low PHB production capacity of methanotrophs continuously cultivated with N_2 with the expression of p MMO were greatly improved in the cyclic NO_3-N_2 cultivation regime, indicating that long-term deficiency of nitrogen sources was detrimental to the activity of methanotrophs expressing p MMO.     

Influence of nonionic surfactant on the solubilization and biodegradation of phenanthrene

Phenanthrene was solubilized in two different nonionic surfactants, Tween80 and Triton X-100. The bioavailability of phenanthrane to the bacteria isolated from the petroleum contaminated soils was studied based on the rotary flasks experiments. The results showed that the concentration of nonionic surfactants above the critical micelle concentration (CMC) can increase the solubility of phenanthrene in water andwere innoxious to the phenanthrene-degrading bacteria; phenanthrene solubilized in the micelles of Tween80 was bioavailable and biodegradable. The research demonstrated the potential of surfactant-enhanced bioremediation of soils contaminated by hydrophobic organic comoounds( HOCs).     

Nitrification characteristics of nitrobacteria immobilized in waterborne polyurethane in wastewater of corn-based ethanol fuel production

A technology to achieve stable and high ammonia nitrogen removal rates for corn distillery wastewater (ethanol fuel production) treatment has been designed. The characteristics of nitrifying bacteria entrapped in a waterborne polyurethane (WPU) gel carrier were evaluated after acclimation. In the acclimation period, nitrification rates of WPU-immobilized nitrobacteria were monitored and polymerase chain reaction (PCR) was also carried out to investigate the change in ammonium-oxidizing bacteria. The results showed that the pellet nitrification rates increased from 21 to 228 mg-N/(L-pellet. hr) and the quantity of the ammonia oxidation bacteria increased substantially during the acclimation. A continuous ammonia removal experiment with the anaerobic pond effluent of a distillery wastewater system was conducted with immobilized nitrifying bacteria for 30 days using an 80 L airlift reactor with pellets at a fill ratio of 15% (V/V). Under the conditions of 75 mg/L influent ammonia, hydraulic retention time (HRT) of 3.7--5.6 hr, and dissolved oxygen (DO) of 4 mg/L, the effluent ammonia concentration was lower than 10 mg/L and the ammonia removal efficiency was 90%. While the highest ammonia removal rate, 162 mg-N/(L-pellet.hr), was observed when the HRT was 1.3 hr.     

Long-term impacts of graphene oxide and Ag nanoparticles on anammox process: Performance, microbial community and toxic mechanism

The increasing application of engineered nanoparticles (NPs) has posed an emerging challenge to constructed wetland wastewater treatment. The performance, microbial community and toxic mechanism of anammox-based unplanted subsurface-flow constructed wetlands (USFCWs) were investigated under the long-term exposure of different graphene oxides (GOs) and Ag NP concentrations. Results showed that the addition of GO could promote TN removal, manifesting as function anammox bacteria C. Anammoxoglobus having a relative high abundance, for GO did not cause significant damage to the cell integrity though there was an increase in ROS concentrations. TN removal would not be obviously affected under exposure of 1?mg/L Ag NPs, for the function gene related to cell biogenesis and repair was up-regulated; while the addition of 10?mg/L Ag NPs would have an inhibiting effect on TN removal in the USFCWs, for the disappearance of some species having anammox ability. Key enzymes of anammox process (NIR and HDH) decreased to some extent under GO and Ag NP exposure, and function gene of defense mechanisms had an increase trend in samples.     

High nitrogen removal from wastewater with several new aerobic bacteria isolated from diverse ecosystems

Introduction The presence of nitrogenous substance has attracted attention because of the role of nitrogen in eutrophication of receiving waters. Denitrification is the ability of bacteria to use nitrogen oxides (NO3- and NO2-) as electron acceptors produ…     

Sequential use of ultraviolet light and chlorine for reclaimed water disinfection

Several disinfection processes of ultraviolet (UV), chlorine or UV followed by chlorine were investigated in municipal wastewater according to the inactivation of Escherichia coli, Shigella dysenteriae and toxicity formation. The UV inactivation of the tested pathogenic bacteria was not affected by the quality of water. It was found that the inactivated bacteria were obviously reactivated after one day in dark. Fluorescent light irradiation increased the bacteria repair. The increase of UV dosage could cause more damage to bacteria to inhibit bacteria self-repair. No photoreactivation was detected when the UV dose was up to 80 mJ/cm² for E. coli DH5αup, and 23 mJ/cm² for S. dysenteriae. Nevertheless, sequential use of 8 mJ/cm² of UV and low concentration of chlorine (1.5 mg/L) could effectively inhibit the photoreactivation and inactivate E. coli below the detection limits within seven days. Compared to chlorination alone, the sequential disinfection decreased the genotoxicity of treated wastewater, especially for the sample with high NH₃-N concentration.     

Quantifying public health benefits of environmental strategy of PM2.5 air quality management in Beijing–Tianjin–Hebei region, China

In 2013,China issued "Air Pollution Prevention and Control Action Plan(Action Plan)" to improve air quality.To assess the benefits of this program in Beijing-Tianjin-Hebei(BTH)region,where the density of population and emissions vary greatly,we simulated the air quality benefit based on Ben MAP to satisfy the Action Plan.In this study,we estimate PM_(2.5) concentration using Voronoi spatial interpolation method on a grid with a spatial resolution of 1 × 1 km~2.Combined with the exposure-response function between PM_(2.5) concentration and health endpoints,health effects of PM_(2.5) exposure are analyzed.The economic loss is assessed by using the willingness to pay(WTP) method and human capital(HC) method.When the PM_(2.5) concentration falls by 25% in BTH and reached 60 μg/m~3 in Beijing,the avoiding deaths will be in the range of 3175 to 14051 based on different functions each year.Of the estimated mortality attributable to all causes,3117 annual deaths were due to lung cancer,1924 – 6318 annual deaths were due to cardiovascular,and343 – 1697 annual deaths were due to respiratory.Based on WTP,the estimated monetary values for the avoided cases of all cause mortality,cardiovascular mortality,respiratory mortality and lung cancer ranged from 1110 to 29632,673 to 13325,120 to 3579,1091 to 6574 million yuan,respectively.Based on HC,the corresponding values for the avoided cases of these four mortalities were 267 to 1178,161 to 529,29 to 143 and 261 million yuan,respectively.     

Natural continuous influent nitrifier immigration effects on nitrification and the microbial community of activated sludge systems

Two sequencing batch reactors (SBRs) were operated for 100 days under aerobic conditions, with one being fed with unsterilized municipal wastewater (USBR), and the other fed with sterilized municipal wastewater (SSBR). Respirometric assays and fluorescence in situ hybridization (FISH) results show that active nitrifiers were present in the unsterilized influent municipal wastewater. The maximum ammonia utilization rate (AUR) and nitrite utilization rate (NUR) of the unsterilized influent were 0.32 ± 0.12 mg NH4+-N/(L·hr) and 0.71 ± 0.18 mg NO2?-N/(L·hr). Based on the maximum utilization rates, the estimated seeding intensity for the ammonia oxidizing bacteria (AOB) and nitrite oxidizing bacteria (NOB) of the USBR was 0.08 g AOB/(g AOB·day) and 0.20 g NOB/(g NOB·day) respectively. The fraction of nitrifiers/total bacteria in the influent was 5.35% ± 2.1%, the dominant AOB was Nitrosomonas spp., Nitrosococcus mobilis hybridizated with Nsm156, and the dominant NOB was Nitrospira hybridizated with Ntspa662. The influent nitrifiers potentially seeded the activated sludge of the bioreactor and hence demonstrated a mitigation of the acclimatization times and instability during start-up and early operation. The AUR and NUR in the USBR was 15% and 13% higher than the SSBR respectively during the stable stage, FISH results showed that nitrifiers population especially the Nitrospira in the USBR was higher than that in the SSBR. These results indicate that the natural continuous immigration of nitrifiers from municipal influent streams may have some repercussions on the modeling and design of bioreactors.     

Quantification of viable bacteria in wastewater treatment plants by using propidium monoazide combined with quantitative PCR (PMA-qPCR)

The detection of viable bacteria in wastewater treatment plants(WWTPs) is very important for public health, as WWTPs are a medium with a high potential for waterborne disease transmission. The aim of this study was to use propidium monoazide(PMA) combined with the quantitative polymerase chain reaction(PMA-qPCR) to selectively detect and quantify viable bacteria cells in full-scale WWTPs in China. PMA was added to the concentrated WWTP samples at a final concentration of 100 μmol/L and the samples were incubated in the dark for 5 min, and then lighted for 4 min prior to DNA extraction and qPCR with specific primers for Escherichia coli and Enterococci, respectively. The results showed that PMA treatment removed more than 99% of DNA from non-viable cells in all the WWTP samples, while matrices in sludge samples markedly reduced the effectiveness of PMA treatment. Compared to qPCR, PMA-qPCR results were similar and highly linearly correlated to those obtained by culture assay, indicating that DNA from non-viable cells present in WWTP samples can be eliminated by PMA treatment, and that PMA-qPCR is a reliable method for detection of viable bacteria in environmental samples. This study demonstrated that PMA-qPCR is a rapid and selective detection method for viable bacteria in WWTP samples, and that WWTPs have an obvious function in removing both viable and non-viable bacteria. The results proved that PMA-qPCR is a promising detection method that has a high potential for application as a complementary method to the standard culture-based method in the future.     

Effects of sulfate-reducing bacteria on methylmercury at the sediment–water interface

Sediment cores(containing sediment and overlying water) from Baihua Reservoir(SW China)were cultured under different redox conditions with different microbial activities, to understand the effects of sulfate-reducing bacteria(SRB) on mercury(Hg) methylation at sediment–water interfaces. Concentrations of dissolved methyl mercury(DMe Hg) in the overlying water of the control cores with bioactivity maintained(BAC) and cores with only sulfate-reducing bacteria inhibited(SRBI) and bacteria fully inhibited(BACI) were measured at the anaerobic stage followed by the aerobic stage. For the BAC and SRBI cores, DMe Hg concentrations in waters were much higher at the anaerobic stage than those at the aerobic stage, and they were negatively correlated to the dissolved oxygen concentrations(r =- 0.5311 and r =- 0.4977 for BAC and SRBI, respectively). The water DMe Hg concentrations of the SRBI cores were 50% lower than those of the BAC cores, indicating that the SRB is of great importance in Hg methylation in sediment–water systems, but there should be other microbes such as iron-reducing bacteria and those containing specific gene cluster(hgc AB), besides SRB,causing Hg methylation in the sediment–water system.     

Phosphorus utilization and microbial community in response to lead/iron addition to a waterlogged soil

Constructed wetlands have emerged as a viable option for helping to solve a wide range of water quality problems. However, heavy metals adsorbed by substrates would decrease the growth of plants, impair the functions of wetlands and eventually result in a failure of contaminant removal. Typha latifolia L., tolerant to heavy metals, has been widely used for phytoremediation of Pb/Zn mine tailings under waterlogged conditions. This study examined e ects of iron as ferrous sulfate (100 and 500 mg/kg) and lead as lead nitrate (0, 100, 500 and 1000 mg/kg) on phosphorus utilization and microbial community structure in a constructed wetland. Wetland plants (T. latifolia) were grown for 8 weeks in rhizobags filled with a paddy soil under waterlogged conditions. The results showed that both the amount of iron plaque on the roots and phosphorus adsorbed on the plaque decreased with the amount of lead addition. When the ratio of added iron to lead was 1:1, phosphorus utilized by plants was the maximum. Total amount of phospholipids fatty acids (PLFAs) was 23%–59% higher in the rhizosphere soil than in bulk soil. The relative abundance of Gram-negative bacteria, aerobic bacteria, and methane oxidizing bacteria was also higher in the rhizosphere soil than in bulk soil, but opposite was observed for other bacteria and fungi. Based on cluster analysis, microbial communities were mostly controlled by the addition of ferrous sulfate and lead nitrate in rhizosphere and bulk soil, respectively.