Flow cytometry (FCM) has been widely used in multi-parametric assessment of cells in various research fields, especially in environmental sciences. This study detected the metabolic activity of Escherichia coli and Staphylococcus aureus by using an FCM method based on 5-cyano-2,3-ditolyltetrazolium chloride (CTC); the accuracy of this method was enhanced by adding SYTO 9 and 10%R2A broth. The disinfection effects of chlorine, chloramine, and UV were subsequently evaluated by FCM methods. Chlorine demonstrated stronger and faster destructive effects on cytomembrane than chloramine, and nucleic acids decomposed afterwards. The metabolic activity of the bacteria persisted after the cytomembranewas damaged as detected using CTC. Low-pressure (LP) UV or medium-pressure (MP) UV treatments exerted no significant effects on membrane permeability. The metabolic activity of the bacteria decreased with increasing UV dosage, and MP-UV was a stronger inhibitor of metabolic activity than LP-UV. Furthermore, the membrane of Gram-positive S. aureus was more resistant to chlorine/chloramine than that of Gram-negative E. coli. In addition, S. aureus showed higher resistance to UV irradiation than E. coli.
A laboratory scale up-flow anaerobic sludge bed (UASB) bioreactor fed with synthetic wastewater was operated with simultaneous methanogenesis and denitrification (SMD) granules for 235 days with a gradient decrease of C/N. Molecular cloning, qRT-PCR and T-RFLP were applied to study the methanogenic community structures in SMD granules and their changes in response to changing influent C/N. The results indicate that when C/N was 20:1, the methane production rate was fastest, and Methanosaetaceae and Methanobacteriaceae were the primary methanogens within the Archaea. The richness and evenness of methanogenic bacteria was best with the highest T-RFLP diversity index of 1.627 in the six granular sludge samples. When C/N was reduced from 20:1 to 5:1, the methanogenic activity of SMD granules decreased gradually, and the relative quantities of methanogens decreased from 36.5% to 10.9%. The abundance of Methanosaetaceae in Archaea increased from 64.5% to 84.2%, while that of Methanobacteriaceae decreased from 18.6% to 11.8%, and the richness and evenness of methanogens decreased along with the T-RFLP diversity index to 1.155, suggesting that the community structure reflected the succession to an unstable condition represented by high nitrate concentrations.
Nutrients and water play an important role in microalgae cultivation. Using wastewater as a culture medium is a promising alternative to recycle nutrients and water, and for further developing microalgae-based products. In the present study, two species of microalgae, Chlorella sp. (high ammonia nitrogen tolerance) and Spirulina platensis (S. platensis, high growth rate), were cultured by using poultry wastewater through a two-stage cultivation system for algal biomass production. Ultrafiltration (UF) or centrifuge was used to harvest Chlorella sp. from the first cultivation stage and to recycle culture medium for S. platensis growth in the second cultivation stage. Results showed the two-stage cultivation system produced high microalgae biomass including 0.39 g·L–1Chlorella sp. and 3.45 g·L–1S. platensis in the first-stage and second-stage, respectively. In addition, the removal efficiencies of NH4+ reached 19% and almost 100% in the first and the second stage, respectively. Total phosphorus (TP) removal reached 17% and 83%, and total organic carbon (TOC) removal reached 55% and 72% in the first and the second stage, respectively. UF and centrifuge can recycle 96.8% and 100% water, respectively. This study provides a new method for the combined of pure microalgae cultivation and wastewater treatment with culture medium recycling.
We assessed the contamination levels of Mn, Zn, Cr, Cu, Ni, Pb, As and Hg and the risks posed by these potentially harmful elements in top-soils around a municipal solid waste incinerator (MSWI).We collected 20 soil samples, with an average pH of 8.1, and another fly ash sample emitted from the MSWI to investigate the concentrations of these elements in soils. We determined the concentrations of these elements by inductively coupled plasma–optical emission spectrometer (ICP-OES), except for Hg, which we measured by AF-610B atomic fluorescence spectrometer (AFS). We assessed the risks of these elements through the use of geoaccumulation index (Igeo), potential ecological risk index (RI), hazard quotient (HQi) and cancer risk (Riski). The results showed that concentrations of potentially harmful elements in soil were influenced by the wind direction, and the concentrations of most elements were higher in the area northwest of the MSWI, compared with the area southeast of the incinerator, with the exception of As; these results were in accordance with those results acquired from our contour maps. According to the Igeo values, some soil samples were clearly polluted by Hg emissions. However, the health risk assessment indicated that the concentrations of Hg and other elements in soil did not pose non-carcinogenic risks to the local populations. This was also the case for the carcinogenic risks posed by As, Cr, and Ni. The carcinogenic risk posed by As was higher, in the range 6.49 × 10–6–9.58 × 10–6, but this was still considered to be an acceptable level of risk.
Polycyclic aromatic hydrocarbons (PAHs) often occur in oil-contaminated soil, coke wastewater and domestic sludge; however, associated PAH degraders in these environments are not clear. Here we evaluated phenanthrene degradation potential in the mixed samples of above environments, and obtained a methanogenic community with different microbial profile compared to those from sediments. Phenanthrene was efficiently degraded (1.26 mg/L/d) and nonstoichiometric amount of methane was produced simultaneously. 16S rRNA gene sequencing demonstrated that bacterial populations were mainly associated with Comamonadaceae Nocardiaceae and Thermodesulfobiaceae, and that methanogenic archaea groups were dominated by Methanobacterium and Methanothermobacter. Substances such as hexane, hexadecane, benzene and glucose showed the most positive effects on phenanthrene degradation. Substrate utilization tests indicated that this culture could not utilize other PAHs. These analyses could offer us some suggestions on the putative phenanthrene-degrading microbes in such environments, and might help us develop strategies for the removal of PAHs from contaminated soil and sludge.
Post-treatment impacts of a novel combined hydrogen peroxide (H2O2) oxidation and WOx/ZrO2 catalysis used for the removal of 1,4-dioxane and chlorinated volatile organic compound (CVOC) contaminants were investigated in soil and groundwater microbial community. This treatment train removed ~90% 1,4-dioxane regardless of initial concentrations of 1,4-dioxane and CVOCs. The Illumina Miseq platform and bioinformatics were used to study the changes to microbial community structure. This approach determined that dynamic shifts of microbiomes were associated with conditions specific to treatments as well as 1,4-dioxane and CVOCs mixtures. The biodiversity was observed to decrease only after oxidation under conditions that included high levels of 1,4-dioxane and CVOCs, but increased when 1,4-dioxane was present without CVOCs. WOx/ZrO2 catalysis reduced biodiversity across all conditions. Taxonomic classification demonstrated oxidative tolerance for members of the genera Massilia and Rhodococcus, while catalyst tolerance was observed for members of the genera Sphingomonas and Devosia. Linear discriminant analysis effect size was a useful statistical tool to highlight representative microbes, while the multidimensional analysis elucidated the separation of microbiomes under the low 1,4-dioxane-only condition from all other conditions containing CVOCs, as well as the differences of microbial population among original, post-oxidation, and post-catalysis states. The results of this study enhance our understanding of microbial community responses to a promising chemical treatment train, and the metagenomic analysis will help practitioners predict the microbial community status during the post-treatment period, which may have consequences for long-term management strategies that include additional biodegradation treatment or natural attenuation.
Under the Stockholm Convention on Persistent Organic Pollutants (POPs), China is required not only to reduce polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans (PCDD/PCDF) but also unintentionally produced polychlorinated biphenyls (PCB), hexachlorobenzene (HCB) and pentachlorobenzene (PeCBz). A baseline of the sources in China that generate these unintentional POPs is needed for both research and regulation purposes. In this paper, we have compiled production data of potential sources in China and assessed them in five-year intervals from 2000 to 2015. Most of these activities experienced changes from rapid growth to slow growth. Measured data for PCB, HCB and PeCBz in samples collected from potential sources in China were reviewed. Most information was associated to thermal processes with high potential of emission, including waste incineration and ferrous and non-ferrous metal production. In addition, high levels of PCB, HCB and PeCBz were found as impurities in a few chlorinated products or as by-products in solvent production, which suggested organochlorine industry might be important sources. Finally, based on the studies reviewed, recommendations for future actions in research and policy as well as a few regulatory issues in China are discussed.
Various pretreatments methods including sonication and grinding were performed on red seaweed Gelidium amansii for the subsequent extraction of agarose. The agarose products are usually extracted from agar powder products from seaweeds. In this study, the agarose was extracted using a direct polyethylene glycol (PEG) method without the need to first process the agar from seaweed. The agar extract was frozen then thawed and mixed directly with PEG solution to precipitate the agarose. The quality of agarose obtained was evaluated through physico-chemical properties analysis which includes spectral technique (FTIR), melting and boiling point, gel strength and sulfate content. These properties were compared with a non-pretreated sample and it was found that the addition of pretreatment steps improved the quality of agarose but gave a slightly lower yield. The gel strength of pretreated samples was much higher and the sulfate content was lower compared to non-pretreated samples. The best pretreatment method was sonication which gave gel strength of 742 g cm-2 and sulfate content of 0.63%. The extraction of agarose can be further improved with the use of different neutralizing agents. Pretreating the seaweed shows potential in improving the quality of agarose from seaweed and can be applied for future extraction of the agarose.
Biofilm is an effective simultaneous denitrification and in situ sludge reduction system, and the characteristics of different biofilm carrier have important implications for biofilm growth and in situ sludge reduction. In this study, the performance and mechanism of in situ sludge reduction were compared between FSC-SBBR and SC-SBBR with constructed by composite floating spherical carriers (FSC) and multi-faceted polyethylene suspension carriers (SC), respectively. The variation of EPS concentration indicated that the biofilm formation of FSC was faster than SC. Compared with SCSBBR, the FSC-SBBR yielded 0.16 g MLSS/g COD, almost 27.27% less sludge. The average removal rates of COD and NH4+-N were 93.39% and 96.66%, respectively, which were 5.21% and 1.43% higher than the average removal rate of SC-SBBR. Investigation of the mechanisms of sludge reduction revealed that, energy uncoupling metabolism and sludge decay were the main factors for sludge reduction inducing 43.13% and 49.65% less sludge, respectively, in FSC-SBBR. EEM fluorescence spectroscopy and SUVA analysis showed that the hydrolytic capacity of biofilm attached in FSC was stronger than those of SC, and the hydrolysis of EPS released more DOM contributed to lysis-cryptic growth metabolism. In additional, Bacteroidetes and Mizugakiibacter associated with sludge reduction were the dominant phylum and genus in FCS-SBBR. Thus, the effect of simultaneous in situ sludge reduction and pollutant removal in FSC-SBBR was better.
Three acid-producing strains, AFB-1, AFB-2 and AFB-3, were isolated during this study, and their roles in anaerobic digestion of waste activated sludge (WAS) were evaluated. Data of 16S rRNA method showed that AFB-1 and AFB-2 were Bacillus coagulans, and AFB-3 was Escherichia coli. The removal in terms of volatile solids (VS) and total chemical oxygen demand (TCOD) was maximized at 42.7% and 44.7% by inoculating Bacillus coagulans AFB-1. Besides, the optimal inoculum concentration of Bacillus coagulans AFB-1 was 30% (v/v). Solubilization degree experiments indicated that solubilization ratios (SR) of WAS reached 20.8%±2.2%, 17.7%±1.48%, and 11.1%±1.53%. Volatile fatty acids (VFAs) concentrations and compositions were also explored with a gas chromatograph. The results showed that VFAs improved by 98.5%, 53.0% and 11.6% than those of the control, respectively. Biochemical methane potential (BMP) experiments revealed that biogas production increased by 90.7% and 75.3% when inoculating with Bacillus coagulans AFB-1 and AFB-2. These results confirmed that the isolated acid-producing bacteria, especially Bacillus coagulans, was a good candidate for anaerobic digestion of WAS.
The aim of this study is to analyze the effect of salinity on polycyclic aromatic hydrocarbons (PAHs) biodegradation, community structure and naphthalene dioxygenase gene (ndo) diversity of a halophilic bacterial consortium with the denaturing gradient gel electrophoresis (DGGE) approach. The consortium was developed from oil-contaminated saline soil after enrichment for six times, using phenanthrene as the substrate. The prominent species in the bacterial consortium at all salinities were identified as halophilic bacteria Halomonas, Alcanivorax, Marinobacter, Idiomarina, Martelella and uncultured bacteria. The predominant microbes gradually changed associating with the saline concentration fluctuations ranging from 0.1% to 25% (w/v). Two ndo alpha subunits were dominant at salinities ranging from 0.1% to 20%, while not been clearly detected at 25% salinity. Consistently, the biodegradation occurred at salinities ranging from 0.1% to 20%, while no at 25% salinity, suggesting the two ndo genes played an important role in the degradation. The phylogenetic analysis revealed that both of the two ndo alpha subunits were related to the classic nah-like gene from Pseudomonas stutzeri AN10 and Pseudomonas aeruginosa PaK1, while one with identity of about 82% and the other one with identity of 90% at amino acid sequence level. We concluded that salinity greatly affected halophilic bacterial community structure and also the functional genes which were more related to biodegradation.
Ibuprofen (IBU) is widely used in the world as anti-inflammatory drug, which posed health risk to the environment. A bacterium capable of degrading IBU was isolated from activated sludge in a sewage treatment plant. According to its morphological, physiologic, and biochemical characteristics, as well as 16S rRNA sequence analysis, the strain was identified as Serratia marcescens BL1 (BL1). Degradation of IBU required the presence of primary substrate. After a five-day cultivation with yeast powder at 30°C and pH 7, the highest degradation (93.47%±2.37%) was achieved. The process of BL1 degrading IBU followed first-order reaction kinetics. The BL1 strain was applied to a small biological aerated filter (BAF) device to form a biofilm with activated sludge. IBU removal by the BAF was consistent with the results of static tests. The removal of IBU was 32.01% to 44.04% higher than for a BAF without BL1. The indigenous bacterial community was able to effectively remove CODMn (permanganate index) and ammonia nitrogen in the presence of BL1.
A spent fluid catalytic cracking (FCC) catalyst containing lanthanum (La) was used as a novel adsorbent for phosphorus (P) in simulated wastewater. The experiments were conducted in a batch system to optimize the operation variables, including pH, calcination temperature, shaking time, solid-liquid ratio, and reaction temperature under three initial P-concentrations (C0 = 0.5, 1.0, and 5.0 mg/L). Orthogonal analysis was used to determine that the initial P-concentration was the most important parameter for P removal. The P-removal rate exceeded 99% and the spent FCC catalyst was more suitable for use in low P-concentration wastewater (C0 <5.0 mg/L). Isotherms, thermodynamics and dynamics of adsorption are used to analyze the mechanism of phosphorus removal. The results show that the adsorption is an endothermic reaction with high affinity and poor reversibility, which indicates a low risk of second releasing of phosphate. Moreover, chemical and physical adsorption coexist in this adsorption process with LaPO4 and KH2PO4 formed on the spent FCC catalyst as the adsorption product. These results demonstrate that the spent FCC catalyst containing La is a potential adsorbent for P-removal from wastewater, which allows recycling of the spent FCC catalyst to improve the quality of water body.
Sulfamethoxazole (SMX) and trimethoprim (TMP) are two critical sulfonamide antibiotics with enhanced persistency that are commonly found in wastewater treatment plants. Recently, more scholars have showed interests in how SMX and TMP antibiotics are biodegraded, which is seldom reported previously. Novel artificial composite soil treatment systems were designed to allow biodegradation to effectively remove adsorbed SMX and TMP from the surface of clay ceramsites. A synergy between sorption and biodegradation improves the removal of SMX and TMP. One highly efficient SMX and TMP degrading bacteria strain, Bacillus subtilis, was isolated from column reactors. In the removal process, this bacteria degrade SMX and TMP to NH4+, and then further convert NH4+ to NO3– in a continuous process. Microbial adaptation time was longer for SMX degradation than for TMP, and SMX was also able to be degraded in aerobic conditions. Importantly, the artificial composite soil treatment system is suitable for application in practical engineering.
The feasibility of using Phragmites australis-JS45 system in removing nitrobenzene from sediments was conducted. However, it was observed that nitrobenzene degraded rapidly and was removed completely within 20 days in native sediments, raising the possibility that indigenous microorganisms may play important roles in nitrobenzene degradation. Consequently, this study aimed to verify this possibility and investigate the potential nitrobenzene degraders among indigenous microorganisms in sediments. The abundance of inoculated strain JS45 and indigenous bacteria in sediments was quantified using real-time polymerase chain reaction. Furthermore, community structure of the indigenous bacteria was analyzed through high throughput sequencing based on Illumina MiSeq platform. The results showed that indigenous bacteria in native sediments were abundant, approximately 1014 CFU/g dry weight, which is about six orders of magnitude higher than that in fertile soils. In addition, the levels of indigenous Proteobacteria (Acinetobacter, Comamonadaceae_ uncultured, Pseudomonas, and Thauera) and Firmicutes (Clostridium, Sporacetigenium, Fusibacter, Youngiibacter, and Trichococcus) increased significantly during nitrobenzene removal. Their quantities sharply decreased after nitrobenzene was removed completely, except for Pseudomonas and Thauera. Based on the results, it can be concluded that indigenous microorganisms including Proteobacteria and Firmicutes can have great potential for removing nitrobenzene from sediments. Although P. australis - JS45 system was set up in an attempt to eliminate nitrobenzene from sediments, and the system did not meet the expectation. The findings still provide valuable information on enhancing nitrobenzene removal by optimizing the sediment conditions for better growth of indigenous Proteobacteria and Firmicutes.
To reveal the distribution characteristics of phytoplankton and the main influence factors under different conditions in the urban rivers, the investigations were conducted during autumn and winter 2014 in Changzhou City, East China. 178 taxa of phytoplankton belonging to 28 functional assemblages were identified. In autumn, the phytoplankton community compositions have high similarity for enhanced hydrological connectivity. The chlorophytes and diatoms (prevailing functional groups C, F, J, P), together with euglenoids (W1), showed high proportions of biomass in the main rivers and connected rivers. It was related to the well mixed eutrophic conditions. The phytoplankton community exhibited spatiotemporal heterogeneity in winter. Affected by the low water level and temperature, the free-living phytoflagellates (X2) replaced groups F and J in the main rivers. Phytoplankton productivity was the highest in the Tongji River. Chlorophytes Dictyosphaerium ehrenbergianum and Chlamydobotrys stellata had an overwhelming superiority during the winter bloom. They were significantly correlated with ammonium, total phosphorus and biochemical oxygen demand. Affected by tail water supply, the diatoms (MP) and euglenoids (W1) dominated in a beheaded river. The multivariate analyses based on the phytoplankton functional groups helped to evaluate the relationships and variations between the urban rivers. The redundancy analysis (RDA) results showed that nitrate nitrogen, water temperature, total nitrogen and total suspended solids were the main influence factors on the phytoplankton community. Except MP, the prevailing groups all showed significant negative correlations with nitrate nitrogen. Availability and utilization of dissolved inorganic nitrogen and hydrodynamic conditions affected the phytoplankton distribution.
Trimethylolpropane (TMP) wastewater is one of the most toxic petrochemical wastewater. Toxicants with high concentrations in TMP wastewater often inhibit the activity of microorganisms associated with biological treatment processes. The hydrolysis acidification process (HAP) is widely used to pretreat petrochemical wastewater. However, the effects of HAP on the reduction of wastewater toxicity and the relevant underlying mechanisms have rarely been reported. In this study, an HAP reactor was operated for 240 days, fed with actual TMP wastewater diluted by tap water in varying ratios. The toxicity of TMP wastewater was assessed with the inhibition ratio of oxygen uptake rate. When the organic loading rates were lower than 7.5 kg COD/m3/d, the toxicity of TMP wastewater was completely eliminated. When the actual TMP wastewater was directly fed into the reactor, the toxicity of TMP wastewater decreased from 100% to 34.9%. According to the results of gas chromatographymass spectrometry analysis, four main toxicants contained in TMP wastewater, namely, formaldehyde, 2-ethylacrolein, TMP and 2-ethylhexanol, were all significantly removed, with removal efficiencies of 93.42%, 95.42%, 72.85% and 98.94%, respectively. Compared with the removal efficiency of CODCr, the reduction rate of toxicity is markedly higher by HAP. In addition, the change of microbial community in the HAP reactor, along the operation period, was studied. The results revealed that, compared with the seed sludge, Firmicutes became the dominant phylum (abundance increased from 0.51% to 57.08%), followed by Proteobacteria and Bacteroidetes (abundance increased from 59.75% to 25.99% and from 4.70% to 8.39%, respectively).
A bioretention unit (BRU) or cell is a green infrastructure practice that is widely used as a low impact development (LID) technique for urban stormwater management. Bioretention is considered a good fit for use in China’s sponge city construction projects. However, studies on bioretention design, which incorporates site-specific environmental and social-economic conditions in China are still very much needed. In this study, an experimental BRU, consisted of two cells planted with Turf grass and Buxus sinica,was tested with eighteen synthesized storm events. Three levels (high, median, low) of flows and concentrations of pollutants (TN, TP and COD) were fed to the BRU and the performance of which was examined. The results showed that the BRU not only delayed and lowered the peak flows but also removed TN, TP and COD in various ways and to different extents. Under the high, medium and low inflow rate conditions, the outflow peaks were delayed for at least 13 minutes and lowered at least 52%. The two cells stored a maximum of 231 mm and 265 mm for turf grass and Buxus sinica, respectively. For both cells the total depth available for storage was 1,220 mm, including a maximum 110 mm deep ponding area. The largest infiltrate rate was 206 mm/h for both cells with different plants. For the eighteen events, TP and COD were removed at least 60% and 42% by mean concentration, and 65% and 49% by total load, respectively. In the reservoir layer, the efficiency ratio of removal of TN, TP and COD were 52%, 8% and 38%, respectively, within 5 days after runoff events stopped. Furthermore, the engineering implication of the hydrological and water quality performances in sponge city construction projects is discussed.
Effect of different carbon sources on purification performance and change of microbial community structure in a novel A2N-MBR process were investigated. The results showed that when fed with acetate, propionate or acetate and propionate mixed (1:1) as carbon sources, the effluent COD, NH4+-N, TN and TP were lower than 30, 5, 15 and 0.5 mg?L–1, respectively. However, taken glucose as carbon source, the TP concentration of effluent reached 2.6 mg?L–1. Process analysis found that the amount of anaerobic phosphorus release would be the key factor to determine the above effectiveness. The acetate was beneficial to the growth of Candidatus Accumulibacter associated with biological phosphorus removal, which was the main cause of high efficiency phosphorus removal in this system. In addition, it could eliminate the Candidatus Competibacter associated with glycogen-accumulating organisms and guarantee high efficiency phosphorus uptake of phosphorus accumulating organisms in the system with acetate as carbon source.
Petroleum hydrocarbons, mainly consisting of n-alkanes and polycyclic aromatic hydrocarbons (PAHs), are considered as priority pollutants and biohazards in the environment, eventually affecting the ecosystem and human health. Though many previous studies have investigated the change of bacterial community and alkane degraders during the degradation of petroleum hydrocarbons, there is still lack of understanding on the impacts of soil alkane contamination level. In the present study, microcosms with different n-alkane contamination (1%, 3% and 5%) were set up and our results indicated a complete alkane degradation after 30 and 50 days in 1%- and 3%-alkane treatments, respectively. In all the treatments, alkanes with medium-chain length (C11-C14) were preferentially degraded by soil microbes, followed by C27-alkane in 3% and 5% treatments. Alkane contamination level slightly altered soil bacterial community, and the main change was the presence and abundance of dominant alkane degraders. Thermogemmatisporaceae, Gemmataceae and Thermodesulfovibrionaceae were highly related to the degradation of C14- and C27-alkanes in 5% treatment, but linked to alkanes with medium-chain (C11-C18) in 1% treatment and C21-alkane in 3% treatment, respectively. Additionally, we compared the abundance of three alkane-monooxygenase genes, e.g., alk_A, alk_P and alk_R. The abundance of alk_R gene was highest in soils, and alk_P gene was more correlated with alkane degradation efficiency, especially in 5% treatment. Our results suggested that alkane contamination level showed non-negligible effects on soil bacterial communities to some extents, and particularly shaped alkane degraders and degrading genes significantly. This study provides a better understanding on the response of alkane degraders and bacterial communities to soil alkane concentrations, which affects their biodegradation process.
