Efficient nitrogen removal pathways and corresponding microbial evidence in tidal flow constructed wetlands for saline water treatment.

The artificial tidal wetlands ecosystem was believed to be a useful device in treating saline water, and it played a significant part in global nitrogen cycles. However, limited information is available on nitrogen-cycling pathways and related contributions to nitrogen loss in tidal flow constructed wetlands (TF-CWs) for saline water treatment. This study operated seven experimental tidal flow constructed wetlands to remove nitrogen from saline water at salinities of 0-30‰. Stable and high NH4+-N removal efficiency (∼90.3%) was achieved, compared to 4.8-93.4% and 23.5-88.4% for nitrate and total nitrogen (TN), respectively. Microbial analyses revealed the simultaneous occurrence of anaerobic ammonium oxidation (anammox), dissimilatory nitrate reduction to ammonium (DNRA), nitrification and denitrification, contributing to nitrogen (N) loss from the mesocosms. The absolute abundances were 5.54 × 103-8.35 × 107 (nitrogen functional genes) and 5.21 × 107-7.99 × 109 copies/g (16S rRNA), while the related genera abundances ranged from 1.81% to 10.47% (nitrate reduction) and from 0.29% to 0.97% (nitrification), respectively. Quantitative response relationships showed ammonium transformation were controlled by nxrA, hzsB and amoA, and nitrate removal by nxrA, nosZ and narG. Collectively, TN transformation were determined by narG, nosZ, qnorB, nirS and hzsB through denitrification and anammox pathways. The proportion of nitrogen assimilation by plants was 6.9-23.4%. In summary, these findings would advance our understanding of quantitative molecular mechanisms in TF-CW mesocosms for treating nitrogen pollution that caused algal blooms in estuarine/coastal ecosystems worldwide.

Molecular analysis of microbial nitrogen transformation and removal potential in the plant rhizosphere of artificial tidal wetlands across salinity gradients.

This study explored the microbial nitrogen transformation and removal potential in the plant rhizosphere of seven artificial tidal wetlands under different salinity gradients (0-30‰). Molecular biological and stable isotopic analyses revealed the existence of simultaneous anammox (anaerobic ammonium oxidation), nitrification, DNRA (dissimilatory nitrate reduction to ammonium) and denitrification processes, contributing to nitrogen loss in rhizosphere soil. The microbial abundances were 2.87 × 103-9.12 × 108 (nitrogen functional genes) and 1.24 × 108-8.43 × 109 copies/g (16S rRNA gene), and the relative abundances of dissimilatory nitrate reduction and nitrification genera ranged from 6.75% to 24.41% and from 0.77% to 1.81%, respectively. The bacterial 16S rRNA high-throughput sequencing indicated that Bacillus, Zobellella and Paracoccus had obvious effects on nitrogen removal by heterotrophic nitrifying/aerobic denitrifying process (HN-AD), and autotrophic nitrification (Nitrosomonas, Nitrospira and Nitrospina), conventional denitrification (Bradyrhizobium, Burkholderia and Flavobacterium), anammox (Candidatus Brocadia and Candidatus Scalindua) and DNRA (Clostridium, Desulfovibrio and Photobacterium) organisms co-existed with HN-AD bacteria. The potential activities of DNRA, nitrification, anammox and denitrification were 1.23-9.23, 400.03-755.91, 3.12-35.24 and 30.51-300.04 nmolN2·g-1·d-1, respectively. The denitrification process contributed to 73.59-88.65% of NOx- reduction, compared to 0.71-13.20% and 8.20-15.42% via DNRA and anammox, as 83.83-90.74% of N2 production was conducted by denitrification, with the rest through anammox. Meanwhile, the nitrification pathway accounted for 95.28-99.23% of NH4+ oxidation, with the rest completed by anammox bacteria. Collectively, these findings improved our understanding on global nitrogen cycles, and provided a new idea for the removal of contaminants in saline water treatment.

Application of light-weight filtration media in an anoxic biofilter for nitrate removal from micro-polluted surface water.

The research investigated nitrate removal from micro-polluted surface water by the single-stage process of anoxic biofilter using light-weight polystyrene beads as filtration media. In this study, sodium acetate was used as an external carbon source and the nitrate removal efficiency under different regimes of hydraulic loading rate (HLR), water temperature, and C/N ratio was studied. In addition, the effect of backwash on denitrification efficiency was investigated. The results show that the biofilter achieved a high nitrate removal efficiency in 2 weeks at water temperatures ranging between 22 and 25 °C at a C/N ratio (COD:NO3(-)-N) of 6:1. Besides, the average removal efficiency of nitrate at HLRs of 5.66, 7.07 and 8.49 m(3) m(-2) h(-1) were 87.5, 87.3 and 87.1%, respectively. The average removal efficiency of nitrate nitrogen was 13.9% at a HLR of 5.66 m(3) m(-2) h(-1) at water temperatures of 12-14 °C, then it increased to 93.7% when the C/N ratio increased to 10. It suggests that the optimal hydraulic retention time is at water temperatures of 8-10 °C. The water consumption rate of backwash was about 0.2-0.3%, and denitrification efficiency returned to the normal level in 12 h after backwash.

Development of a constructed wetland water treatment system for selenium removal: incorporation of an algal treatment component.

On the basis of the fact that algae have the ability to volatilize substantial quantities of selenium (Se), we investigated the concept of including an algal pretreatment unit into a constructed wetland system for the removal of Se from river water entering the Salton Sea. Of six different algal strains tested, the most effective in terms of Se volatilization and Se removal from the water column was a Chlorella vulgaris strain (designated Cv). Cv removed 96% of Se (supplied as selenate) from the microcosm water column within 72 h, with up to 61% being removed by volatilization to the atmosphere. X-ray absorption spectroscopy revealed that the major forms of Se likely to be accumulated in an algal-wetland system are selenomethionine, a precursor of volatile Se formation, and elemental Se. Our results suggest that the inclusion of an algal pretreatment unit within a constructed wetland water treatment system should not only enhance the efficiency of Se removal but also significantly reduce the risk of the buildup of ecotoxic forms of Se by promoting the biological volatilization of Se.

Development of a constructed wetland water treatment system for selenium removal: use of mesocosms to evaluate design parameters.

The Salton Sea in California is an important habitat for fish and waterfowl. Its ecosystem is threatened due to diminishing water supplies and increasing salinity. An alternative source of water to support species conservation habitat may be obtained from local rivers (e.g., Alamo or New Rivers), provided that a wetland treatment system can be developed to remove selenium (Se), fertilizer nutrients, and other contaminants. Here, we used mesocosms to evaluate a number of potential design options (e.g., plant species selection, sediment composition and arrangement, forced aeration, organic amendments, etc.) to improve the efficiency of Se removal using treatment wetlands. Our results show that, of five different substrate arrangements tested for Se removal, the most efficient was obtained for cattails growing in a substrate of cattail litter overlying sand and peat moss sediment (water column Se was reduced from 15 µg Se/L to <0.1 µg Se/L in 72 h). The addition of organic amendments in the form of alfalfa hay or alfalfa meal was also helpful in lowering Se levels. These results suggest that it may be possible to design constructed wetland water treatment systems capable of reducing Se concentrations in river water to values below 1 µg Se/L.

Bioremediation of selenium-contaminated soil using earthworm Eisenia fetida: Effects of gut bacteria in feces on the soil microbiome.

Selenium (Se) contamination in the soil poses a food safety risk to humans. The present study was to investigate the role of earthworm Eisenia fetida in soil Se remediation. When exposed to selenite at 4 mg Se/kg, E. fetida efficiently concentrated Se in tissues (24.53 mg Se/kg dry weight), however, only accounting for a minor portion of the added Se. Microbial analysis shows 12 out of 15 functional genera became more abundant in the worm-inhabited soil when exposed to Se, suggesting E. fetida contributed to Se remediation mainly by introducing Se-reducing bacteria to the soil via feces, which were dominated by the genera Pseudomonas (∼62.65%) and Aeromonas (∼29.99%), whose abundance was also significantly boosted in the worm-inhabited soil. However, when isolated from worm feces at 200 mg Se/L, Pseudomonas strains only displayed a high tolerance to Se rather than removal capacity. In contrast, among 4 isolated Aeromonas strains, A. caviae rapidly removing 85.74% of the added selenite, mainly through accumulation (67.38%), while A. hydrophila and A. veronii were more effective at volatilizing Se (27.77% and 24.54%, respectively), and A. media performed best by reducing Se by ∼49.00% under anaerobic conditions. Overall, our findings have highlighted the importance of E. fetida as a key contributor of functional bacteria to the soil microbiome, building a strong foundation for the development of an earthworm-soil system for Se bioremediation.

Sulfur transformation in sulfur autotrophic denitrification using thiosulfate as electron donor.

Thiosulfate is frequently used as an energy source and electron donor in autotrophic denitrification (AD) for removing nitrate from wastewater. However, transforming pathways of S2O32- in this process is unclear. Herein, the aim of this study is to explore possible transforming pathways of sulfur compounds in thiosulfate-based AD process. After measuring the variation of NO3-, NO2-, and various sulfur compounds such as S0, SO42-, S2O32-, acid volatile sulfide (AVS), and S2- in the presence and absence of S2O32-, the variation process of S2O32- and the contribution of various sulfur compounds were analyzed. The results indicated that S0, AVS, and S2- were the intermediate products when S2O32- was applied as an electron donor. All S2O32-, S0, AVS, and S2- could act as electron donors in the nitrate removal process with the final products of SO42-. The utilization priority of these four sulfur sources was presumed in the following order: S2- > S2O32- > AVS ≈ S0. Furthermore, sulfur transformation and balance in nitrate removal process was also investigated. This suggests the transforming pathways of sulfur compounds in denitrification process. Nitrogen removal and sulfur conversion process are dependent on the presence of microorganisms in the sludge.

Improvement of nitrogen removal with iron scraps in floating treatment wetlands.

Floating treatment wetland (FTW) in restoration of low C/N ratio wastewater was deemed to a frequently used method. However, the nitrate removal performance in floating beds was limited due to insufficient organic carbon sources. Iron scraps as a potential electron donor was beneficial to the NO3--N reduction. To research the removal performance and mechanism of denitrification in FTW with iron scraps, FTW with Iris pseudacorus was built, and iron scraps were added as an electron donor to improve nitrogen removal efficiency. The batch experimental results demonstrated that the proper mass ratio of iron scraps to NO3--N was 500:1. With iron scraps, the NO3--N removal efficiency of FTW and control system increased significantly to 98.04% and 44.42% respectively in 2 weeks, while there was no obvious influence on the removal of NH4+-N. After adding iron scraps, the proportion of bacteria in the systems related to iron cycle and the relative abundance of nitrifying and denitrifying bacteria have increased obviously. By calculating the nitrogen balance, nitrogen reduction via plant uptake accounted for 8.79%, and the microbial denitrification was the main nitrogen removal pathway in FTW.

Efficient utilization of Iris pseudacorus biomass for nitrogen removal in constructed wetlands: Combining alkali treatment.

Aquatic plant biomass like Iris pseudacorus can be used as electron donor to improve denitrification performance in subsurface constructed wetlands. However, the phenomenon that the nitrogen removal rate declined in the terminal stage restricted the utilization of litters. In terms of this problem, this study investigated the performance of the used biomass through alkali treatment on nitrogen removal and analyzed the effect of alkali treatment on the component and structure of biomass and microbial community. The results showed that the alkali-treated biomass could further enhance the nitrogen removal by nearly 15% compared with used ones. The significant damage of cell walls and compact fibers containing cellulose and lignin through alkali treatment mainly resulted in the improvement of carbon release and nitrogen removal. With the addition of alkali-treated biomass, the richness index of microbes was higher compared with other biomass materials. Furthermore, the abundance of denitrification related genera increased and the abundance of genera for nitrification was maintained. Based on these finds, a mode of a more efficient Iris pseudacorus self-consumed subsurface flow constructed wetlands was designed. In this mode, the effluent total nitrogen could be stabilized below 5 mg L-1 for nine months and the weight of litters could be further cut down by 75%. These findings would contribute to efficient utilization of plant biomass for nitrogen removal enhancement and final residue reduction in the wetlands.

Recycled utilization of Iris pseudacorus in constructed wetlands: Litters self-consumption and nitrogen removal improvement.

Aquatic plants litters from constructed wetlands might become pollutants without proper treatment. Due to its high carbon and low nitrogen contained, Iris pseudacorus litters have potential to be used as carbon source to enhance denitrification process in advanced treatments of secondary effluent from wastewater treatment plants. This study investigated the characteristics of carbon release form Iris pseudacorus litters and its performance on enhancement of nitrogen removal. The batch experiment showed that the organic carbon release process can be simulated by combining dissolution and hydrolysis process, and it was found that dissolved organic matters mainly consisted of 60% sugar and 35% humic acid-like compounds from the neutral detergent solution and hemicellulose of litters. The long-term operation of lab-scale constructed wetlands revealed a high nitrogen removal of 78.81-90.39% in treating the synthetic wastewater treatment plants effluent with the equivalent dosage of 25-150 g litters m-2 d-1. Furthermore, it is possible to establish an Iris pseudacorus self-consumed constructed wetland to reuse all of the litters produced during the operation. These findings can contribute to the understanding of the dynamics of carbon release from Iris pseudacorus litters and recycled utilization of plant biomass in the constructed wetlands.

Selenium uptake, volatilization, and transformation by the cyanobacterium Microcystis aeruginosa and post-treatment of Se-laden biomass.

With a narrow margin between beneficial and toxic effects, selenium (Se) is of great concern due to its increasing level in aquatic environments. The accumulation and transformation of Se by the cyanobacterium Microcystis aeruginosa and effects of nutrients, particularly sulfate, were investigated. The nutrient-deprived cyanobacterium removed water-borne selenate (82.2 ± 0.93%) faster than selenite (58.9 ± 1.77%), with 86.0 ± 1.41% and 77.2 ± 1.00%, respectively, of the Se accumulated in the biomass and the rest volatilized. When supplied with excess nutrients, the Se accumulation and volatilization rates were significantly inhibited, with the removal efficiency dropping to 50.2 ± 2.59% and 7.37 ± 0.93% for selenite and selenate, respectively. When M. aeruginosa was tested with inadequate, appropriate, and adequate levels of sulfate, Se uptake decreased with increasing sulfate concentrations, particularly for selenate (from 34.1 to 4.81%). Using X-ray absorption near-edge structure to speciate biomass Se, selenite and selenate were transformed to organo-Se (87.3-100%), with or without nutrients present, suggesting M. aeruginosa could efficiently reduce Se oxyanions to more bioavailable forms. With increasing sulfate levels (5.0 and 10.0 mg S/L), percentages of SeMet converted from selenite decreased by 28.2-33.0%, with 19.1-33.2% as elemental Se, while organo-Se remained dominant (93.6-95.1%) in selenate-treated M. aeruginosa. Transmission electron microscopy shows structural damage in the cell wall at exposure to selenite (1600 µg Se/L), with the intracellular structure intact. To prevent Se biomagnification along aquatic food chains, the Se-laden biomass was combusted as a post-treatment, leading to a significant reduction in Se content (∼99.2%) and Se bioavailability, with inorganic Se (45.0-70.5%) predominant in the residue.

Biological iron nitrogen cycle in ecological floating bed: Nitrogen removal improvement and nitrous oxide emission reduction.

Ecological floating beds (EFBs) have become a superior method for treating secondary effluent from wastewater treatment plant. However, insufficient electron donor limited its denitrification efficiency. Iron scraps from lathe cutting waste consist of more than 95% iron could be used as electron donors to enhance denitrification. In this study, EFBs with and without iron scraps supplementation (EFB-Fe and EFB, respectively) were conducted to explore the impacts of iron scraps addition on nitrogen removal, nitrous oxide (N2O) emissions and microbial communities. Results showed the total nitrogen (TN) removal in EFB-Fe improved to 79% while that in EFB was 56%. N2O emission was 0-6.20 mg m-2 d-1 (EFB-Fe) and 1.74-15.2 mg m-2 d-1 (EFB). Iron scraps could not only improve nitrogen removal efficiency, but also reduce N2O emissions. In addition, high-throughput sequencing analysis revealed that adding iron scraps could improve the sum of denitrification related genera, among which Novosphingobium accounted for the highest proportion (6.75% of PFe1, 4.24% of PFe2, 3.18% of PFe3). Iron-oxidizing bacteria and iron-respiring bacteria associated with and nitrate reducing bacteria mainly concentrated on the surface of iron scraps. Principal co-ordinates analysis (PCoA) indicated that iron scraps were the key factor affecting microbial community composition. The mechanism of iron scraps enhanced nitrogen removal was realized by enhanced biological denitrification process. Iron release dynamic from iron scraps was detected in bench-scale experiment and the electron transfer mechanism was that Fe0 transferred electrons directly to NO3--N, and biological iron nitrogen cycle occurred in EFB-Fe without secondary pollution.

Remediation of selenium-contaminated soil through combined use of earthworm Eisenia fetida and organic materials.

Selenium (Se)-polluted soils pose serious threats to terrestrial ecosystems through food chains. This study evaluated the use of earthworm Eisenia fetida and organic materials for Se remediation. The greater mortality (6.7%) and weight loss (28.6%) were observed for earthworms exposed to selenate than selenite at 20 mg Se Kg-1 over 21 d, while selenate was taken up 2.5-fold faster than selenite. Compared with peat moss, Se accumulation increased by 119% in selenite-exposed earthworms supplied with cow manure. Earthworm activity caused greater reduction in selenite (17.4%), with little impact on selenate-treated soil. X-ray absorption spectroscopy speciation analysis shows 87-91% of tissue Se was transformed to organo-Se, i.e., SeMet and SeCys, in earthworms exposed to either selenite or selenate, posing great risks to their predators. The study also found selenium increased bacterial diversity in earthworm casts, while greater relative abundances (~37.57%) of functional genera were obtained for selenite. Over 24 h, two bacteria strains, Bacillus cereus and Aeromonas encheleia, isolated from casts, rapidly reduced selenite by ~94%, compared to ~25% for selenate. Elemental Se was present only in strains (~27%), casts (~11%) and worm-inhabited soil (~2.7%) of selenite treatments, suggesting earthworm gut microbiota could buffer earthworms and other soil fauna from selenite toxicity.

Nitrogen removal through collaborative microbial pathways in tidal flow constructed wetlands.

Constructed wetlands are efficient in removing nitrogen from water; however, little is known about nitrogen-cycling pathways for nitrogen loss from tidal flow constructed wetlands. This study conducted molecular and stable isotopic analyses to investigate potential dissimilatory nitrate reduction to ammonium (DNRA), denitrification, nitrification, anaerobic ammonium oxidation (anammox), and their contributions to nitrogen removal by two tidal wetland mesocosms, PA (planted with Phragmites australis) and NP (unplanted), designated to treat Yangtze River Estuary water. Our results show the mesocosms removed ~22.6% of TN from nitrate-dominated river water (1.19 mg·L-1), with better performance obtained in PA than that in NP, which was consistent with the molecular and stable isotopic data. The potential activities of DNRA, anammox, denitrification and nitrification varied between 0.6 and 1.6, 4.6-37.3, 36.4-305.7, and 463.7-945.9 nmol N2 g-1 dry soil d-1, respectively, with higher values obtained in PA than NP. Nitrification accounted for 94.3-99.4% of NH4+ oxidation, with the rest through anammox. Denitrification contributed to 77.9-90.3% of NOx- reduction, compared to 9.2-21.6% and 0.5-1.5% via anammox and DNRA, respectively; 78.4-90.9% of N2 was produced through denitrification, with the rest via anammox. Pearson correlation analyses suggest NH4+ was the major factor regulating nitrification, while NO3- played an important role in the competition between denitrification and DNRA, and NO2- was a key restrictive factor for anammox. Overall, this study reveals the importance of nitrification, denitrification, anammox and DNRA in nitrogen removal, providing new insight into the nitrogen-cycling mechanisms in natural/artificial tidal wetlands.

Salinity-driven nitrogen removal and its quantitative molecular mechanisms in artificial tidal wetlands.

The present study investigated the performance in nitrogen removal and associated nitrogen transformation processes in seven mesocosms fed with saline water (0‰ to 30‰) to simulate tidal flow constructed wetlands (TF CWs). The highly effective and steady removal of NH4+-N (84.74% averagely) was obtained at various salinities, while the rates varied from 6.34% to 89.19% and 22.54% to 87.48% for NO3--N and total nitrogen (TN), respectively. Overall, nitrogen removal efficiencies were greater at lower salinities. Molecular biological analyses verified the co-occurrence of dissimilatory nitrate reduction to ammonium (DNRA), denitrification, anaerobic ammonium oxidation (anammox) and nitrification in the mesocosms, reportedly contributing to nitrogen removal in TF CWs. The absolute copy numbers of nitrogen functional genes and total bacterial 16S rRNA were 2.54 × 103-7.35 × 107 and 3.21 × 107-7.82 × 109 copies g-1 dg (dry gravel), respectively, with the dominant phyla, i.e., Chloroflexi, Proteobacteria, Actinobacteriota, Cyanobacteria, and Firmicutes, accounting for over 80% of the sequences. The relative abundances of the genera related to nitrification and dissimilatory nitrate reduction processes, i.e., denitrification, anammox and DNRA, varied from 0.16% to 0.89% and from 3.66% to 11.59%, respectively, while quantitative relationships confirmed NH4+-N transformation rate was jointly controlled by amoA, hzsB, nxrA and nrfA, and NO3--N removal rate by nirS, nosZ, narG, qnorB and nxrA. These findings may shed light on quantitative molecular mechanisms for nitrogen removal in TF CWs for the saline water treatment, providing a sustainable solution to nitrogen pollution problem in the estuary ecosystem.

Se transformation and removal by a cattail litter treatment system inoculated with sulfur-based denitrification sludge: Role of the microbial community composition under various temperature and aeration conditions.

With a narrow margin between deficiency and toxicity, rising levels of selenium (Se) are threatening aquatic ecosystems. To investigate the role of microorganisms in Se bioremediation, a cattail litter system inoculated with the sulfur-based denitrification sludge was conducted. The results show the litter, as a carrier and nutrient source for bacteria, efficiently removed Se by ~ 97.0% during a 12-d treatment with water circulating. As the major removal pathways, immobilization rates of selenite were ~ 2.9-fold higher than selenate, and the volatilization, contributing to ~ 87.7% of the total Se removal, was significantly correlated with temperature (positively) and oxidation-reduction potential (ORP; negatively). Using X-ray absorption spectroscopy to speciate litter-borne Se, more Se0 formed without aeration due to abundant Se-reducing bacteria, among which Azospira and Azospirillum were highly related to the removal of both Se oxyanions, while Desulfovibrio, Azoarcus, Sulfurospirillum, Thauera, Geobacter, Clostridium, and Pediococcus were the major contributors to selenate removal. Overall, our study suggests microbial Se metabolism in the litter system was significantly affected by temperature and ORP, which could be manipulated to enhance Se removal efficiency and the transformation of selenate/selenite into low toxic Se0 and volatile Se, reducing risks posed by the residual Se in the system.

Enhancement of a constructed wetland water treatment system for selenium removal.

Selenium (Se) is essential to most animals, whereas the gap between necessity and toxicity is narrow. Our previous work showed constructed wetlands were a promising solution to Se contamination in aquatic habitats. This study further examined effects of organic amendments and hydrologic regimes on Se removal by constructed wetlands. Our results suggest the removal efficiency exceeded 94% within 8 days for the systems with moderate and low organic carbon contents in the substrate, as a 98% removal of Se was obtained in three weeks for the system subjected to the 2-day wet/dry cycle. To mimic field wetlands, a litter layer was added to the cattail treatment system, which reduced waterborne Se much more rapidly than control, achieving a 77% removal of Se within 4 days. XAS results show all sediment Se was transformed to Se0 in the presence of litter, as SeMet (47%) dominated the Se adsorbed by the litter. The findings indicate the Se removal capacity of a constructed wetland would improve over time, especially via Se volatilization into the atmosphere and Se stabilization in the sediment with litter accumulating at the surface. Another mesocosm experiment showed the cattail floating system effectively removed Se, particularly selenate, by 99% in 48 h. To confirm that high performance, seven constructed wetland types were set up for comparison. The results show the cattail floating system was most effective in Se removal (93-100% at around 35 °C in summer and 51-100% at about 5 °C in winter). More research is needed to test the floating system under more field conditions and investigate the biomagnification and biotransformation of the removed Se along food chains. Seven constructed wetland types were set up for comparison.

Interactive effects of earthworm Eisenia fetida and bean plant Phaseolus vulgaris L on the fate of soil selenium.

Selenium (Se) is an essential micronutrient for animals with a narrow margin between essentiality and toxicity. Se toxicity is largely related to inorganic forms of Se in soil, i.e., selenite and selenate that enter food chains through plant uptake, threatening higher trophic level organisms. This experiment investigated effects of earthworm activity on Se bioavailability in soil and the subsequent plant uptake, using earthworm Eisenia fetida and bean plant Phaseolus vulgaris L, both exposed to either selenite or selenate at 1 or 4 mg Se kg-1 for 16 weeks. Plants took up selenate (up to 221-fold) faster than selenite, with up to 84% of the Se rapidly transported to shoots. In the presence of earthworms, Se accumulation obviously increased for selenate-supplied plants, leading to an up to 4% increase in Se translocation factor for all treatments except for 1 mg kg-1 selenite treatment. Earthworms also concentrated Se faster in tissues (up to 274 mg kg-1 DW) at exposure to selenate. For Se toxicity, Se speciation analysis was conducted on the plants and earthworms using XAS. Compared to worm-free treatments, the percentage of organo-Se, i.e., SeMet and CysSeSeCys, increased in beans (up to 34%) in the presence of earthworms for selenate, while the elemental Se portion was significantly reduced or absent, opposite to the results for selenite. Surprisingly, elemental Se (up to 65%) dominated earthworms, regardless of the form of Se supplied. In conclusion, earthworms clearly enhanced Se uptake and translocation in plants, leading to elevated Se levels in shoots. To prevent resulting hazards to humans and other animals, caution should be taken while consuming the shoots, particularly beans, harvested from the Se contaminated soil where earthworm activity is high. Finally, the significant reduction in soil Se suggests phytoextraction of Se from the soil could be improved using earthworms as an aid to plants.

Application of a novel two-stage biofiltration system for simulated brackish aquaculture wastewater treatment.

Biofiltration is one kind of common technology used for treating micro-polluted brackish aquaculture wastewater. Based on the characteristics of actual water quality, a novel two-stage biofiltration system was set up to reduce potential nutrient pollution brought by the frequent exchange of water in brackish pond aquaculture. Zeolite was selected as filtration media for the first stage and pyrite mixed with a small amount of sulfur for the second stage. Apart from the adsorption of nutrients exerted by these natural minerals, biofilm played a leading role in nutrient removal. The surface and internal pore of zeolite-sheltered nitrifiers and sulfur-containing compounds enhanced autotrophic denitrification. It was found that ammonia adsorption capacity of zeolite was reduced by nearly 58% when salinity was increased to 1.5%, while phosphate adsorption capacity of pyrite was hardly influenced and systematic hydraulic retention time (HRT) of 24 h was proven appropriate, 9.6 h and 14.4 h for the two stages, respectively. Meanwhile, removal efficiency of 96.5% for NH4+-N and 92.1% for total inorganic nitrogen (TIN) was achieved under this condition. The analysis of microbial community of biofilm indicated that dominant genera responsible for nitritation and nitration on the surface of zeolite were Nitrosomonas and Nitrospira, respectively. Dominant genera responsible for autotrophic denitrification on the surface of pyrite and sulfur were both Thiobacillus. In addition, Ferritrophicum, related to the iron-oxidizing bacterium, also coexisted due to biological oxidation of pyrite. Long-term operation verified applicability and stability of this two-stage biofiltration system for brackish aquaculture wastewater purification.

Trophic transfer and biotransformation of selenium in the mosquito (Aedes albopictus) and interactive effects with hexavalent chromium.

As an essential micronutrient for animals with a narrow range between essentiality and toxicity, selenium (Se) usually coexists with chromium (Cr) in contaminated aquatic environments. This study investigated effects of three diets (Microcystis aeruginosa, Chlorella vulgaris and biofilms) exposed to Se or/and Cr on Aedes albopictus as a vector for the aquatic-terrestrial transfer of Se and Cr. Se(IV)-exposed mosquitoes concentrated Se up to 66-fold faster than Se(VI)-exposed ones, corresponding to the greater Se enrichment in Se(IV)-treated diets. Analysis using synchrotron-based X-ray absorption spectroscopy (XAS) showed that Se(0) (61.9-74.6%) dominated Se(VI)-exposed mosquitoes except for the C. vulgaris-fed larvae (organo-Se, 94.0%), while organo-Se accounted for 93.3-100.0% in Se(IV)-exposed mosquitoes. Cr accumulation in larvae (56.40-87.24 µg Cr/g DW) or adults (19.41-50.77 µg Cr/g DW) was not significantly different among all Cr(VI) treatments, despite varying diet Cr levels. With Cr(0) being dominant (57.7-94.0%), Cr(VI)-exposed mosquitoes posed little threat to predators. Although mosquitoes exposed to Se or Cr had shorter wings, adults supplied with C. vulgaris or biofilms co-exposed to Se(VI) and Cr(VI) had wings significantly (1.1-1.2 fold) longer than Se(VI) only exposed ones. Overall, our study reveals the role of Ae. albopictus in transferring waterborne Se and Cr from the contaminated aquatic ecosystem to the terrestrial ecosystem with the resulting eco-risks to wildlife in both ecosystems.