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In this study, we isolated two bacterial strains, P2 (Pseudomonas geniculata ATCC 19374, P. geniculata ATCC 19374) and B1 (Bacillus cereus EC3, B. cereus EC3) from pig farm soil, which were mixed to construct a novel bacterial mixture (P2:B1 = 1:1). We studied the heterotrophic nitrification capablity and growth characteristics of bacterial mixture in heterotrophic nitrification medium (HNM). And the factors affecting the nitrogen removal of the bacterial mixture such as C/N ratio, NH4+-N load and inoculation size of the bacterial mixture were investigated in this study. Furthermore, we used the bacterial mixture to treat livestock wastewater to remove NH4+-N, TN and COD. The results showed that the optimal C/N ratio and inoculation size for the bacterial mixture were 12 and 5%, respectively. It is worth to mention that the bacterial mixture shows excellent NH4+-N removal ability under low C/N ratio, indicating that the bacterial mixture had the potential to be applied to wastewater with C/N lower than 8. When the initial NH4+-N concentration was 500 mg/L, the NH4+-N removal efficiency exceeded 90% with a 5% inoculation size of the bacterial mixture after 72 h. When the single bacteria and bacterial mixture were employed to treat livestock wastewater, the removal efficiencies of bacterial mixture on NH4+-N, TN and COD within 72 h were 70.06%, 58.12% and 81.48%, respectively, which were higher than that of single bacteria. The bacterial mixture has potential application value in enhancing the ability of reactors and constructed wetlands to remove pollutants from livestock wastewater and developing a novel and efficient biological nitrogen removal technology. Our current research provides a research basis for the application of the bacterial mixture in the future.

A heterotrophic nitrifying bacterium was isolated from the activated sludge and identified as Pseudomonas stutzeri GEP-01. Strain GEP-01 exhibited an efficient heterotrophic nitrification capability and a high nitrogen utilization rate, 48 mg/L NH4+-N was removed after culturing for 24 h without NO2−-N or NO3−-N accumulation, and 64.7% of the NH4+-N was removed by heterotrophic nitrification. Single-factor experiments indicated that factors such as the carbon source, temperature, NH4+-N load, and inoculum size had significant effects on the ammonium removal efficiency of strain GEP-01. The preferred conditions for heterotrophic nitrification were sodium citrate, 30 °C, 40 mg/L NH4+-N, and 5% inoculum size. When the initial NH4+-N amounts were 100, 200, 500 and 1000 mg/L, the removal rates were approximately 100%, 93%, 90.4%, and 78.9%, respectively, and higher ammonium concentrations require longer culture time. Nitrogen balance demonstrated that 40% of the initial nitrogen was lost, which was probably removed in the form of gas products under optimum culture conditions, and 36.3% of NH4+-N was converted to biomass. When incubated (adding a small amount of sodium citrate as carbon source and no carbon source) in swine wastewater containing 835 mg/L of ammonium, the removal ratio reached 56.3% and 24.8%. Strain GEP-01 has potential applications in the treatment of ammonium-rich wastewater.

The removal efficiency of contaminants in large-scale integrated vertical-flow constructed wetland (IVCW) and vertical-flow constructed wetland (VCW) for wastewater treatment plant (WWTP) tail-water was evaluated, and the microbial community was also investigated in this study. The results for 14 months study period indicated that 40.05% chemical oxygen demand (COD), 45.47% ammonia nitrogen (NH4+-N), 62.55% total phosphorus (TP), 55.53% total nitrogen (TN) and 57.20% total suspended solids (TSS) average removal efficiencies were achieved in the IVCW. There was a poor performance of TN removal in the VCW, with an average removal efficiency of 38.13%. There was no significant seasonal difference in TP removal, and a strong positive correlation between influent TP load and removed load. The high-throughput sequencing analysis revealed that Proteobacteria, Planctomycetes, Bacteroidetes and Acidobacteria were dominant in nature and wetland systems. The relative abundance of nitrifying bacteria, denitrifying bacteria and anammox bacteria confirmed that nitrification, denitrification and anammox may be the main processes for nitrogen removal in the IVCW.

In this study, the evergreen Iris sibirica plant was planted in a constructed wetland during the low-temperature season. Microcosmic subsurface vertical flow constructed wetlands (MVFCWs) were fed with simulated Cd polluted river water for three months under different Cd supply and the seasonal growth changes and Cd uptake ability of Iris sibirica were investigated. The treatment performance of MVFCWs and removal mechanisms and distribution of Cd were evaluated. The amount of rhizospheric microorganisms (bacteria, fungi, and actinomyces) and the enzyme activities (urease, sucrase, alkaline phosphatase and catalase) were also monitored. Iris sibirica proves to be a good accumulator of Cd with the highest accumulation of Cd in below-ground parts of 443.3 mg/kg when exposed to 8 mg/L Cd for three months. Under 1, 2, 4 and 8 mg/L Cd supply in low-temperature seasons, the MVFCWs exhibit average Cd removals of 94.9%, 97.4%, 95.6% and 96.2%, respectively. The predominant removal process of Cd in MVFCWs is substrate adsorption, which accounts for about 50% of Cd removal. Under the same Cd supply, the removal rates of Cd, amount of rhizospheric microbes and activities of enzymes in planted wetland are higher than those in unplanted wetland.

A symbiotic microalgal-bacterial system may be an optional technology for wastewater treatment. In this study, co-immobilized of a bacterium isolated from a municipal wastewater treatment plant ( Pseudomonas putida ) and a microalgae Chlorella vulgaris was used in the study of cell growth and nutrient removal during wastewater treatment under batch and continuous culture conditions. Under batch culture conditions, co-immobilization treatment significantly increased the cell density of C. vulgaris and P. putida compared with other treatments. The co-immobilized treatment also showed higher removal of ammonium, phosphate and COD than any single treatment, indicating that the nutrient uptake capability of C. vulgaris and P. Putida was mutually enhanced mutually. When tested in continuous mode, the treatment with a hydraulic retention time of 24 h at the organic load rate of 1159.2 mg COD L −1 d −1 was most appropriate for wastewater treatment.

Ammonia has been a major reason of macrophyte decline in the water environment, and ammonium ion toxicity should be seen as universal, even in species frequently labeled as "NH4+ specialists". To study the effects of high NH4+-N stress of ammonium ion nitrogen on tolerant submerged macrophytes and investigate the pathways of nitrogen assimilation in different organisms, Myriophyllum aquaticum was selected and treated with various concentrations of ammonium ions at different times. Increasing of ammonium concentration leads to an overall increase in incipient ammonia content in leaves and stems of plants. In middle and later stages, high concentrations of NH4+ ion nitrogen taken up by M. aquaticum decreased, whereas the content of NO3- ion nitrogen increased. Moreover, in M. aquaticum, the activities of the enzymes nitrate reductase, glutamine synthetase and asparagine synthetase changed remarkably in the process of alleviating NH4+ toxicity and deficiency. The results of the present study may support the studies on detoxification of high ammonium ion content in NH4+-tolerant submerged macrophytes and exploration of tissue-specific expression systems.

Enrichment of the hyperaccumulator bank is important for phytoremediation, and studying new hyperaccumulators has become a research hotspot. In this study, cadmium (Cd), the main representative factor of heavy-metal-polluted water, was the research object, and the Cd bioenrichment ability and tolerance of Myriophyllum aquaticum were studied for the first time. The experiment was conducted for 28 days by establishing experimental groups with different Cd concentrations (0, 10, 20, 40, 80, and 160 mg/L). The results show that M. aquaticum is a new Cd hyperaccumulator. There was no notable damage in the 40 mg/L Cd treatment group, and the Cd enrichment ability of M. aquaticum reached 17,970 ± 1020.01 mg/kg, while the bioconcentration factor (BCF) reached 449.25. At the same time, the antioxidant system (superoxide dismutase (SOD) and peroxidase (POD)) and proline (Pro) levels of M. aquaticum maintained normal plant physiology, but there were physiological anomalies in M. aquaticum at high concentrations and under long-term treatment. The results show that M. aquaticum has a high Cd bioenrichment ability and tolerance in water and can be used for phytoremediation of river water polluted by Cd.

Three submerged macrophytes, Ceratophyllum demersum (CD), Myriophyllum spicatum (MS) and Myriophyllum aquaticum (MA), were treated with various concentrations of ammonia for different lengths of time. Ammonium ions (NH4+) in the medium severely inhibited plant growth and led to a reduction in total chlorophyll (chl a and b) in CD and MS. The addition of ammonia significantly decreased the soluble protein content and increased the free amino acid content of CD and MS in treatments with high concentrations of NH4+, but MA showed no significant physiological response. The antioxidant enzyme system of MA was activated, which in turn reduced the peroxidation level in the plant and maintained the plant’s normal physiological activities when the ammonia nitrogen in the culture fluid increased. The study continued to use higher concentrations (25, 50, 100, 200 and 400 mg/L) of ammonium nitrogen to treat and observe the peroxidation level and corresponding enzyme production for this species of MA in vivo to explore its resistance mechanism. The experiments show that MA can normally live for a period of time in a high-ammonia environment of up to 100 mg/L. The results of the present study will assist in studies of the detoxification of high ammonium ion contents in submersed macrophytes and the selection of plants suitable for macrophyte recovery.

The objective of the present study was to determine the effect of total ammonia N concentration and pH on N metabolism of Ceratophyllum demersum and to evaluate stress as a result of inorganic N enrichment in the water column on submerged macrophytes. Carefully controlled pH values distinguished between the effects of un-ionized NH3 and ionized NH4(+). The results showed that the most obvious consequence of ammonia addition was an overall increase in ammonia content and decrease in nitrate content in all tissues of fertilized plants. The activities of nitrate reductase and glutamine synthetase were inhibited by long-term ammonia addition. At the same time, ammonia addition significantly decreased soluble protein content and increased free amino acid content in all treatments. Another clear effect of ammonia addition was a decrease in carbon reserves. Therefore, the authors concluded that increased ammonia availability could affect plant survival and lead to a decline in C. demersum proliferation through a decrease in their carbon reserves. This interaction between N and C metabolism helps to explain changes in benthic vegetation as a result of steadily increasing coastal water eutrophication.

In this study, the tips of Myriophyllum aquaticum (M. aquaticum) plants were planted in open-top plastic bins and treated by simulated wastewater with various ammonium-N concentrations for three weeks. The contents of related carbohydrates and key enzyme activities of carbon metabolism were measured, and the mechanisms of carbon metabolism regulation of the ammonia tolerant plant M. aquaticum under different ammonium-N levels were investigated. The decrease in total nonstructural carbohydrates, soluble sugars, sucrose, fructose, reducing sugar and starch content of M. aquaticum were induced after treatment with ammonium-N during the entire stress process. This finding showed that M. aquaticum consumed a lot of carbohydrates to provide energy during the detoxification process of ammonia nitrogen. Moreover, ammonia-N treatment led to the increase in the activitives of invertase (INV) and sucrose synthase (SS), which contributed to breaking down more sucrose to provide substance and energy for plant cells. Meanwhile, the sucrose phosphate synthase (SPS) activity was also enhanced under stress of high concentrations of ammonium-N, especially on day 21. The result indicated that under high-concentration ammonium-N stress, SPS activity can be significantly stimulated by regulating carbon metabolism of M. aquaticum, thereby accumulating sucrose in the plant body. Taken together, M. aquaticum can regulate the transformation of related carbohydrates in vivo by highly efficient expression of INV, SPS and SS, and effectively regulate the osmotic potential, thereby delaying the toxicity of ammonia nitrogen and improving the resistance to stress. It is very important to study carbon metabolism under ammonia stress to understand the ammonia nitrogen tolerance mechanism of M. aquaticum.

A novel process for sewage sludge bioleaching by mixed Thiobacilli (Acidithiobacillus thiooxidans and Acidithiobacillus ferrooxidans) using a 12-stage, 180 L working volume continuous plug-flow bioreactor, is presented. The objective of the present study was to assess the impact of some parameters on the sludge dewaterability and to improve the sludge dewaterability by optimization of these parameters. The parameters examined were sludge moisture content, nutrients dosage, aeration rate, and the number of reactors. The order of the influence of these factors on sludge dewaterability was found to be sludge moisture content > nutrients dosage > aeration rate > number of reactors. The optimized conditions were: sludge moisture content, 98.0%, nutrients dosage, 9 g/L, aeration rate, 8 m3/h, and 10 reactors. Confirmation experiments conducted under optimum conditions demonstrate the sludge dewaterability to be remarkably improved. After 2 days of bioleaching, the moisture of bioleached sludge cake was reduced to below 60%.

In the present study, carefully controlled pH ranges (7 and 9) were used to distinguish between the effects of un-ionized NH3 and the NH4+ ion. The objective was to find the effect of different total ammonia nitrogen concentrations and pH values on the carbon metabolism of Ceratophyllum demersum. The authors investigated the effects of ammonia on the nonstructural carbohydrate content in shoots of C. demersum. Ammonia treatment decreased the contents of nonstructural carbohydrate, soluble sugar, sucrose, fructose, and starch in leaves. Meanwhile, increasing the pH value exacerbated the decline of the C. demersum nonstructural carbohydrate content. In addition, the activity of invertase was increased during the experiment. These results suggest that ammonia severely inhibits plant growth by disturbing nonstructural carbohydrate content. It has been suggested that ammonia has toxic effects on C. demersum and that the higher the pH in water, the more obvious the physiological responses that C. demersum exhibits. The results of the present study can provide some reference for studying the living conditions of submersed macrophytes under the stress of NH3. Environ Toxicol Chem 2015;34:843–849. © 2014 SETAC

Fluoride can either inhibit or enhance the growth of aquatic macrophytes, depending upon fluoride concentration and exposure time. To investigate fluoride toxicity, the submerged plant Hydrilla verticillata was treated with various concentrations of fluoride (F) (0, 10, 20, and 40 mg/L) for different lengths of time (7, 14, 21, and 28 days). At exposure to 10 mg/L F, the content of chlorophyll, protein, and carbohydrates content increased in leaves of H. verticillata, and the activity of guaiacol peroxidase (POD) and superoxide dismutase (SOD) slightly increased in plants compared with the control. When fluoride concentration increased to 20 mg/L, the toxic effect generated by fluoride led to a reduction of chlorophyll, protein, and carbohydrates in H. verticillata, but the activity of guaiacol peroxidase and SOD and the amount of ascorbic acid (AsA) and glutathione (GSH) were enhanced significantly. After exposure to fluoride at 40 mg/L for a long period, these physiological parameters showed a sharp decrease, and inactivation was observed in H. verticillata. These results suggested that a certain concentration of fluoride induced antioxidant response, and excess fluoride induced metabolism imbalance and oxidative damage in H. verticillata.