Search results for: denitrifying%20bioreactor

Authors: Meenakshi Srivastava, A. K. Mishra

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This study characterizes the metabolic versatility of denitrifying bacterial communities residing in the paddy soil using the GC-MS based Phospholipid Fatty Acid (PLFA) analyses simultaneously with nosZ gene based PCR-DGGE (Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis) and real time Q-PCR analysis. We have analyzed the abundance of nitrous oxide reductase (nosZ) genes, which was subsequently related to soil PLFA profile and DGGE based denitrifier community structure. Soil denitrifying bacterial community comprised majority or dominance of Ochrobactrum sp. following Cupriavidus and uncultured bacteria strains in paddy soil of selected sites. Initially, we have analyzed the abundance of the nitrous oxide reductase gene (nosZ), which was found to be related with PLFA based lipid profile. Chandauli of Eastern UP, India represented greater amount of lipid content (C18-C20) and denitrifier’s diversity. This study suggests the positive co-relation between soil PLFA profiles, DGGE, and Q-PCR data. Thus, a close networking among metabolic abilities and taxonomic composition of soil microbial communities existed, and subsequently, such work at greater extent could be helpful in managing nutrient dynamics as well as microbial dynamics of paddy soil ecosystem.

Keywords: denaturing gradient gel electrophoresis, DGGE, nitrifying and denitrifying bacteria, PLFA, Q-PCR

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Authors: Prangya Ranjan Rout, Puspendu Bhunia, Rajesh Roshan Dash

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In a developing country like India, providing reliable and affordable wastewater treatment facilities in rural areas is a huge challenge. With the aim of enhancing the nutrient removal from rural domestic wastewater while reducing the cost of treatment process, a novel, integrated treatment system consisting of a multistage bio-filter with drop aeration and a post positioned attached growth carbonaceous denitrifying-bioreactor was designed and developed in this work. The bio-filter was packed with ‘dolochar’, a sponge iron industry waste, as an adsorbent mainly for phosphate removal through physiochemical approach. The Denitrifying bio-reactor was packed with many waste organic solid substances (WOSS) as carbon sources and substrates for biomass attachment, mainly to remove nitrate in biological denitrification process. The performance of the modular system, treating real domestic wastewater was monitored for a period of about 60 days and the average removal efficiencies during the period were as follows: phosphate, 97.37%; nitrate, 85.91%, ammonia, 87.85%, with mean final effluent concentration of 0.73, 9.86, and 9.46 mg/L, respectively. The multistage bio-filter played an important role in ammonium oxidation and phosphate adsorption. The multilevel drop aeration with increasing oxygenation, and the special media used, consisting of certain oxides were likely beneficial for nitrification and phosphorus removal, respectively, whereas the nitrate was effectively reduced by biological denitrification in the carbonaceous bioreactor. This treatment system would allow multipurpose reuse of the final effluent. Moreover, the saturated dolochar can be used as nutrient suppliers in agricultural practices and the partially degraded carbonaceous substances can be subjected to composting, and subsequently used as an organic fertilizer. Thus, the system displays immense potential for treating domestic wastewater significantly decreasing the concentrations of nutrients and more importantly, facilitating the conversion of the waste materials into usable ones.

Keywords: nutrient removal, denitrifying bioreactor, multi-stage bio-filter, dolochar, waste organic solid substances

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Authors: A. Jéglot, F. Plauborg, M. K. Schnorr, R. S. Sørensen, L. Elsgaard

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Artificial wetlands such as woodchip bioreactors are efficient tools to remove nitrate from agricultural wastewater with a minimized environmental impact. However, the temperature dependence of the microbiological nitrate removal prevents the woodchip bioreactors from being an efficient system when the water temperature drops below 8℃. To quantify and describe the temperature effects on nitrate removal efficiency, we studied nitrate-reducing enrichments from a woodchip bioreactor in Denmark based on samples collected in Spring and Fall. Growth was quantified as optical density, and nitrate and nitrous oxide concentrations were measured in time-course experiments to compare the growth of the microbial population and the nitrate conversion efficiencies at different temperatures. Ammonia was measured to indicate the importance of dissimilatory nitrate reduction to ammonia (DNRA) in nitrate conversion for the given denitrifying community. The temperature responses observed followed the increasing trend proposed by the Arrhenius equation, indicating higher nitrate removal efficiencies at higher temperatures. However, the growth and the nitrous oxide production observed at low temperature provided evidence of the psychrotolerance of the microbial community under study. The assays conducted showed higher nitrate removal from the microbial community extracted from the woodchip bioreactor at the cold season compared to the ones extracted during the warmer season. This indicated the ability of the bacterial populations in the bioreactor to evolve and adapt to different seasonal temperatures.

Keywords: agricultural waste water treatment, artificial wetland, denitrification, psychrophilic conditions

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Authors: Danish Laidin, Peter Gostomski, Aaron Marshall, Carlo Carere

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Groundwater contamination of nitrate (NO3-) is becoming more prevalent in regions of intensive and extensive agricultural activities. Household nitrate removal involves using ion exchange membranes and reverse osmosis (RO) systems, whereas industrial nitrate removal may use organic carbon substrates (e.g. methanol) for heterotrophic microbial denitrification. However, these approaches both require high capital investment and operating costs. In this study, denitrification was demonstrated using bio-electrochemical systems (BESs) inoculated from sediments and microbial enrichment cultures. The BES reactors were operated continuously as microbial electrolytic cells (MECs) with a poised potential of -0.7V and -1.1V vs Ag/AgCl. Three parallel MECs were inoculated using hydrogen-driven denitrifying enrichments, stream sediments, and biofilm harvested from a denitrifying biotrickling filter, respectively. These reactors were continuously operated for over a year as various operating conditions were investigated to determine the optimal conditions for electroactive denitrification. The mass loading rate of nitrate was varied between 10 – 70 mg NO3-/d, and the maximum observed nitrate removal rate was 22 mg NO3- /(cm2∙d) with a current of 2.1 mA. For volumetric load experiments, the dilution rate of 1 mM NO3- feed was varied between 0.01 – 0.1 hr-1 to achieve a nitrate loading rate similar to the mass loading rate experiments. Under these conditions, the maximum rate of denitrification observed was 15.8 mg NO3- /(cm2∙d) with a current of 1.7mA. Hydrogen (H2) was supplied intermittently to investigate the hydrogenotrophic potential of the denitrifying biofilm electrodes. H2 supplementation at 0.1 mL/min resulted in an increase of nitrate removal from 0.3 mg NO3- /(cm2∙d) to 3.4 mg NO3- /(cm2∙d) in the hydrogenotrophically subcultured reactor but had no impact on the reactors which exhibited direct electron transfer properties. Results from this study depict the denitrification performance of the immobilized biofilm electrodes, either by direct electron transfer or hydrogen-driven denitrification, and the contribution of the planktonic cells present in the growth medium. Other results will include the microbial community analysis via 16s rDNA amplicon sequencing, varying the effect of poising cathodic potential from 0.7V to 1.3V vs Ag/AgCl, investigating the potential of using in-situ electrochemically produced hydrogen for autotrophic denitrification and adjusting the conductivity of the feed solution to mimic groundwater conditions. These findings highlight the overall performance of sediment inoculated MECs in removing nitrate and will be used for the future development of sustainable solutions for the treatment of nitrate polluted groundwater.

Keywords: bio-electrochemical systems, groundwater, electroactive denitrification, microbial electrolytic cell

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Authors: Lina Wu

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The excessive discharge of nitrogen in sewage greatly intensifies the eutrophication of water bodies and threatens water quality. Nitrogen pollution control has become a global concern. The concentration of nitrogen in water is reduced by converting ammonia nitrogen, nitrate nitrogen and nitrite nitrogen into nitrogen-containing gas through biological treatment, physicochemical treatment and oxidation technology. However, some wastewater containing high ammonia nitrogen including landfill leachate, is difficult to be treated by traditional nitrification and denitrification because of its high COD content. The core process of denitrification is that denitrifying bacteria convert nitrous acid produced by nitrification into nitrite under anaerobic conditions. Still, its low-carbon nitrogen does not meet the conditions for denitrification. Many studies have shown that the natural autotrophic anammox bacteria can combine nitrous and ammonia nitrogen without a carbon source through functional genes to achieve total nitrogen removal, which is very suitable for removing nitrogen from leachate. In addition, the process also saves a lot of aeration energy consumption than the traditional nitrogen removal process. Therefore, anammox plays an important role in nitrogen conversion and energy saving. The short-range nitrification and denitrification coupled with anaerobic ammoX ensures total nitrogen removal. It improves the removal efficiency, meeting the needs of society for an ecologically friendly and cost-effective nutrient removal treatment technology. In recent years, research has found that the symbiotic system has more water treatment advantages because this process not only helps to improve the efficiency of wastewater treatment but also allows carbon dioxide reduction and resource recovery. Microalgae use carbon dioxide dissolved in water or released through bacterial respiration to produce oxygen for bacteria through photosynthesis under light, and bacteria, in turn, provide metabolites and inorganic carbon sources for the growth of microalgae, which may lead the algal bacteria symbiotic system save most or all of the aeration energy consumption. It has become a trend to make microalgae and light-avoiding anammox bacteria play synergistic roles by adjusting the light-to-dark ratio. Microalgae in the outer layer of light particles block most of the light and provide cofactors and amino acids to promote nitrogen removal. In particular, myxoccota MYX1 can degrade extracellular proteins produced by microalgae, providing amino acids for the entire bacterial community, which helps anammox bacteria save metabolic energy and adapt to light. As a result, initiating and maintaining the process of combining dominant algae and anaerobic denitrifying bacterial communities has great potential in treating landfill leachate. Chlorella has a brilliant removal effect and can withstand extreme environments in terms of high ammonia nitrogen, high salt and low temperature. It is urgent to study whether the algal mud mixture rich in denitrifying bacteria and chlorella can greatly improve the efficiency of landfill leachate treatment under an anaerobic environment where photosynthesis is stopped. The optimal dilution concentration of simulated landfill leachate can be found by determining the treatment effect of the same batch of bacteria and algae mixtures under different initial ammonia nitrogen concentrations and making a comparison. High-throughput sequencing technology was used to analyze the changes in microbial diversity, related functional genera and functional genes under optimal conditions, providing a theoretical and practical basis for the engineering application of novel bacteria-algae symbiosis system in biogas slurry treatment and resource utilization.

Keywords: nutrient removal and recovery, leachate, anammox, Partial nitrification, Algae-bacteria interaction

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Authors: V. Ivanov, J. Chu, V. Stabnikov

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Low cost, sustainable, and environmentally friendly microbial cements, grouts, polysaccharides and bioplastics are useful in construction and geotechnical engineering. Construction-related biotechnologies are based on activity of different microorganisms: urease-producing, acidogenic, halophilic, alkaliphilic, denitrifying, iron- and sulphate-reducing bacteria, cyanobacteria, algae, microscopic fungi. The bio-related materials and processes can be used for the bioaggregation, soil biogrouting and bioclogging, biocementation, biodesaturation of water-satured soil, bioencapsulation of soft clay, biocoating, and biorepair of the concrete surface. Altogether with the most popular calcium- and urea based biocementation, there are possible and often are more effective such methods of ground improvement as calcium- and magnesium based biocementation, calcium phosphate strengthening of soil, calcium bicarbonate biocementation, and iron- or polysaccharide based bioclogging. The construction-related microbial biotechnologies have a lot of advantages over conventional construction materials and processes.

Keywords: ground improvement, biocementation, biogrouting, microorganisms

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Authors: Rupak Kumar Sarma, Ratul Saikia

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Macrophytes encompass major functional group in eutrophic wetland ecosystems. As a key functional element of freshwater lakes, they play a crucial role in regulating various wetland biogeochemical cycles, as well as maintain the biodiversity at the ecosystem level. The high carbon-rich underground biomass of macrophyte populations may harbour diverse microbial community having significant potential in maintaining different biogeochemical cycles. The present investigation was designed to study the macrophyte-microbe interaction in coupled nitrification and denitrification, considering Deepor Beel Lake (a Ramsar conservation site) of North East India as a model eutrophic system. Highly eutrophic sites of Deepor Beel were selected based on sediment oxygen demand and inorganic phosphorus and nitrogen (P&N) concentration. Sediment redox potential and depth of the lake was chosen as the benchmark for collecting the plant and sediment samples. The average highest depth in winter (January 2016) and summer (July 2016) were recorded as 20ft (6.096m) and 35ft (10.668m) respectively. Both sampling depth and sampling seasons had the distinct effect on variation in macrophyte community composition. Overall, the dominant macrophytic populations in the lake were Nymphaea alba, Hydrilla verticillata, Utricularia flexuosa, Vallisneria spiralis, Najas indica, Monochoria hastaefolia, Trapa bispinosa, Ipomea fistulosa, Hygrorhiza aristata, Polygonum hydropiper, Eichhornia crassipes and Euryale ferox. There was a distinct correlation in the variation of major sediment physicochemical parameters with change in macrophyte community compositions. Quantitative estimation revealed an almost even accumulation of nitrate and nitrite in the sediment samples dominated by the plant species Eichhornia crassipes, Nymphaea alba, Hydrilla verticillata, Vallisneria spiralis, Euryale ferox and Monochoria hastaefolia, which might have signified a stable nitrification and denitrification process in the sites dominated by the selected aquatic plants. This was further examined by a systematic analysis of microbial populations through culture dependent and independent approach. Culture-dependent bacterial community study revealed the higher population of nitrifiers and denitrifiers in the sediment samples dominated by the six macrophyte species. However, culture-independent study with bacterial 16S rDNA V3-V4 metagenome sequencing revealed the overall similar type of bacterial phylum in all the sediment samples collected during the study. Thus, there might be the possibility of uneven distribution of nitrifying and denitrifying molecular markers among the sediment samples collected during the investigation. The diversity and abundance of the nitrifying and denitrifying molecular markers in the sediment samples are under investigation. Thus, the role of different aquatic plant functional types in microorganism mediated nitrogen cycle coupling could be screened out further from the present initial investigation.

Keywords: denitrification, macrophyte, metagenome, microorganism, nitrification

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Authors: V. Ivanov, J. Chu, V. Stabnikov

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Different biotechnological methods for the production of construction materials and for the performance of construction processes in situ are developing within a new scientific discipline of Construction Biotechnology. The aim of this research was to develop and test new biotechnologies and biotechnological grouts for the minimization of the hydraulic conductivity of the fractured rocks and porous soil. This problem is essential to minimize flow rate of groundwater into the construction sites, the tunneling space before and after excavation, inside levies, as well as to stop water seepage from the aquaculture ponds, agricultural channels, radioactive waste or toxic chemicals storage sites, from the landfills or from the soil-polluted sites. The conventional fine or ultrafine cement grouts or chemical grouts have such restrictions as high cost, viscosity, sometime toxicity but the biogrouts, which are based on microbial or enzymatic activities and some not expensive inorganic reagents, could be more suitable in many cases because of lower cost and low or zero toxicity. Due to these advantages, development of biotechnologies for biogrouting is going exponentially. However, most popular at present biogrout, which is based on activity of urease- producing bacteria initiating crystallization of calcium carbonate from calcium salt has such disadvantages as production of toxic ammonium/ammonia and development of high pH. Therefore, the aim of our studies was development and testing of new biogrouts that are environmentally friendly and have low cost suitable for large scale geotechnical, construction, and environmental applications. New microbial biotechnologies have been studied and tested in the sand columns, fissured rock samples, in 1 m3 tank with sand, and in the pack of stone sheets that were the models of the porous soil and fractured rocks. Several biotechnological methods showed positive results: 1) biogrouting using sequential desaturation of sand by injection of denitrifying bacteria and medium following with biocementation using urease-producing bacteria, urea and calcium salt decreased hydraulic conductivity of sand to 2×10-7 ms-1 after 17 days of treatment and consumed almost three times less reagents than conventional calcium-and urea-based biogrouting; 2) biogrouting using slime-producing bacteria decreased hydraulic conductivity of sand to 1x10-6 ms-1 after 15 days of treatment; 3) biogrouting of the rocks with the width of the fissures 65×10-6 m using calcium bicarbonate solution, that was produced from CaCO3 and CO2 under 30 bars pressure, decreased hydraulic conductivity of the fissured rocks to 2×10-7 ms-1 after 5 days of treatment. These bioclogging technologies could have a lot of advantages over conventional construction materials and processes and can be used in geotechnical engineering, agriculture and aquaculture, and for the environmental protection.

Keywords: biocementation, bioclogging, biogrouting, fractured rocks, porous soil, tunneling space

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Authors: Kraiem Khadija, Hamadi Kallali, Naceur Jedidi

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Constructed wetland systems are eco-technology recognized as environmentally friendly and emerging innovative solutions remediation as these systems are cost-effective and sustainable wastewater treatment systems. The performance of these biological system is affected by various factors such as plant, substrate, wastewater type, hydraulic loading rate, hydraulic retention time, water depth, and operation mood. The objective of this study was to to assess the alters of plant species on pollutants reduction and enrichment of anammox and nitrifing denitrifing bacteria in a modified vertical flow (VFCW) constructed wetland. This tests were carried out using three modified vertical constructed wetlands with a surface of 0.23 m² and depth 80 cm. It was a saturated vertical constructed wetland at the bottom. The saturation zone is maintained by the siphon structure at the outlet. The VFCW (₁) system was unplanted, VFCW (₂) planted with Typha angustofolia, and VFCW(₃) planted with Phragmites australis. The experimental units were fed with domestic wastewater and were operated by batch mode during 8 months at an average hydraulic loading rate around 20 cm day− 1. The operation cycle was two days feeding and five days rest. Results indicated that plants presence improved the removal efficiency; the removal rates of organic matter (85.1–90.9%; COD and 81.8–88.9%; BOD5), nitrogen (54.2–73%; NTK and 66–77%; NH4 -N) were higher by 10.7–30.1% compared to the unplanted vertical constructed wetland. On the other hand, the plant species had no significant effect on removal efficiency of COD, The removal of COD was similar in VFCW (₂) and VFCW (₃) (p > 0.05), attaining average removal efficiencies of 88.7% and 85.2%, respectively. Whereas it had a significant effect on NTK removal (p > 0.05), with an average removal rate of 72% versus 51% for VFCW (₂) and VFCW (₃), respectively. Among the three sets of vertical flow constructed wetlands, the VFCW(₂) removed the highest percent of total streptococcus, fecal streptococcus total coliforms, fecal coliforms, E. coli as 59, 62, 52, 63, and 58%, respectively. The presence and the plant species alters the community composition and abundance of the bacteria. The abundance of bacteria in the planted wetland was much higher than that in the unplanted one. VFCW(₃) had the highest relative abundance of nitrifying bacteria such as Nitrosospira (18%), Nitrosospira (12%), and Nitrobacter (8%). Whereas the vertical constructed wetland planted with typha had larger number of denitrifying species, with relative abundances of Aeromonas (13%), Paracoccus (11%), Thauera (7%), and Thiobacillus (6%). However, the abundance of nitrifying bacteria was very lower in this system than VFCW(₂). Interestingly, the presence of Thypha angustofolia species favored the enrichment of anammox bacteria compared to unplanted system and system planted with phragmites australis. The results showed that the middle layer had the most accumulation of anammox bacteria, which the anaerobic condition is better and the root system is moderate. Vegetation has several characteristics that make it an essential component of wetlands, but its exact effects are complex and debated.

Keywords: wastawater, constructed wetland, anammox, removal

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Authors: A. Chávez, A. Rodríguez, F. Pinzón

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Society is concerned about measures of environmental, economic and social impacts generated in the solid waste disposal. These places of confinement, also known as landfills, are locations where problems of pollution and damage to human health are reduced. They are technically designed and operated, using engineering principles, storing the residue in a small area, compact it to reduce volume and covering them with soil layers. Problems preventing liquid (leachate) and gases produced by the decomposition of organic matter. Despite planning and site selection for disposal, monitoring and control of selected processes, remains the dilemma of the leachate as extreme concentration of pollutants, devastating soil, flora and fauna; aggressive processes requiring priority attention. A biological technology is the activated sludge system, used for tributaries with high pollutant loads. Since transforms biodegradable dissolved and particulate matter into CO2, H2O and sludge; transform suspended and no Settleable solids; change nutrients as nitrogen and phosphorous; and degrades heavy metals. The microorganisms that remove organic matter in the processes are in generally facultative heterotrophic bacteria, forming heterogeneous populations. Is possible to find unicellular fungi, algae, protozoa and rotifers, that process the organic carbon source and oxygen, as well as the nitrogen and phosphorus because are vital for cell synthesis. The mixture of the substrate, in this case sludge leachate, molasses and wastewater is maintained ventilated by mechanical aeration diffusers. Considering as the biological processes work to remove dissolved material (< 45 microns), generating biomass, easily obtained by decantation processes. The design consists of an artificial support and aeration pumps, favoring develop microorganisms (denitrifying) using oxygen (O) with nitrate, resulting in nitrogen (N) in the gas phase. Thus, avoiding negative effects of the presence of ammonia or phosphorus. Overall the activated sludge system includes about 8 hours of hydraulic retention time, which does not prevent the demand for nitrification, which occurs on average in a value of MLSS 3,000 mg/L. The extended aeration works with times greater than 24 hours detention; with ratio of organic load/biomass inventory under 0.1; and average stay time (sludge age) more than 8 days. This project developed a pilot system with sludge leachate from Doña Juana landfill - RSDJ –, located in Bogota, Colombia, where they will be subjected to a process of activated sludge and extended aeration through a sequential Bach reactor - SBR, to be dump in hydric sources, avoiding ecological collapse. The system worked with a dwell time of 8 days, 30 L capacity, mainly by removing values of BOD and COD above 90%, with initial data of 1720 mg/L and 6500 mg/L respectively. Motivating the deliberate nitrification is expected to be possible commercial use diffused aeration systems for sludge leachate from landfills.

Keywords: sludge, landfill, leachate, SBR

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