Metal biosorption in lignocellulosic biofuel biorefinery effluent: an initial step towards sustainability of water resources

Biosorption of metals by microorganisms is a promising technology to remove accumulated non-process elements in highly recycled biorefinery process water. Removal of these elements would enable greater water reuse and reduce the environmental impact of effluent discharge. A model lignocellulosic ethanol biorefinery wastewater was created based on pulp mill effluent. This generated a wastewater with an environmentally realistic high loading of dissolved natural organic matter (900?mg/l), a potentially important factor influencing metal biosorption. Analysis of feedstock and pulp mill effluent indicated that Mn and Zn are likely to be problematic in highly recycled lignocellulosic ethanol biorefinery process water. Therefore, the growth of several bacteria and fungi from existing collections, and some isolated from pulp mill effluent were tested in the model wastewater spiked with Mn and Zn (0.2?mM). Wastewater isolates grew the best in the wastewater. Metal uptake varied by species and was much greater for Zn than Mn. A bacterium, Novosphingobium nitrogenifigens Y88(T), removed the most metal per unit biomass, 35 and 17?mg?Mn/g. No other organism tested decreased the Mn concentration. A yeast, Candida tropicalis, produced the most biomass and removed the most total metal (38?% of Zn), while uptake per unit biomass was 24?mg?Zn/g. These results indicate that microorganisms can remove significant amounts of metals in wastewater with high concentrations of dissolved natural organic matter. Metal sorption by autochthonous microorganisms in an anaerobic bioreactor may be able to extend water reuse and therefore lower the water consumption of future biorefineries.     

Cadmium and mercury removal from non-point source wastewater by a hybrid bioreactor

The purpose of this study was to remove cadmium (Cd) and mercury (Hg) from non-point source wastewater by a hybrid bioreactor consisting of two different processes (anaerobic–anoxic–aerobic and photoautotrophic). The results showed that the bioreactor could concurrently culture heterotrophic and autotrophic microorganisms, and removed Cd and Hg from the wastewater successfully. The average removal efficiencies were 79% and 66%, respectively for Cd and Hg. The relationship between Cd removal rate and biofilm mass was observed to be significant (p < 0.05) during different seasons. The Hg removal was mainly due to the bioaccumulation in macrophytes via a photoautotrophic process. Due to the increase of the bacterial diversity under the rejuvenated conditions modulated by the hybrid bioreactor, the growth conditions of the native bacterial habitat were improved. The results demonstrate that the environmentally benign, easily-deployed, sludge free and cost-effective hybrid bioreactor can efficiently remove Cd and Hg from non-point source wastewater.     

The removal of nutrients from non-point source wastewater by a hybrid bioreactor

The aim of this project was to establish an economical and environmentally benign biotechnology for removing nutrients from non-point source wastewater. The proposal involves a hybrid bioreactor comprised of sequential anaerobic, anoxic and aerobic (A²/O) processes and an eco-ditch being constructed and applied in a suburban area, Kunming, south-western China, where wastewater was discharged from an industrial park and suburban communities. The results show that the hybrid bioreactor fosters heterotrophic and autotrophic microorganisms. When the hydraulic load is 200 m³ per day with the running mode in 12 h cycles, the removal efficiencies of the nutrients were 81% for TP, 74% for TDP, 82% for TN, 79% for NO₃-N and 86% for NH₄-N. The improved bacterial community structure and bacterial habitats further implied enhanced water quality and indicates that the easily-deployed, affordable and environmentally-friendly hybrid bioreactor is a promising bio-measure for removing high loadings of nutrients from non-point source wastewater.     

PPCPs removal by aerobic granular sludge membrane bioreactor

An aerobic granular sludge membrane bioreactor (GMBR) was applied to the treatment of pharmaceutical and personal care products (PPCPs) wastewater. The influence of granular sludge on five antibiotic and antiphlogistic PPCPs wastewater and the removal effect of methyl alcohol and conventional organic matter were investigated while constantly reducing the density of inflow organic matter. The results showed that the sludge granulation process in the system was rapid but unstable, and that the system exhibits a dissolution–reunion dynamic equilibrium. The reactor demonstrated varying removal effects of PPCPs on different objects. The use of a GMBR was more effective for the removal of prednisolone, naproxen, and ibuprofen; the first two drugs were lower the average removal rate of which reached 98.46 and 84.02 %, respectively; whereas the average removal rate of ibuprofen was 63.32 %. By contrast, the GMBR has an insignificant degradation effect on antibiotics such as amoxicillin, indicating that such antibiotic medicine is not easily degraded by microorganisms, which plays different roles in system operation. Because of the different chemical structures and characteristics of drugs that result in various degradation behavior. During the GMBR granulation process, the value of mixed liquor volatility suspended solids (MLVSS) gradually increases from 1.5 to 4.1 g/L during the GMBR granulation process, and the removal rate of COD_Cr reaches up to 87.98 %. After reducing the density of organic matter is reduced, the removal rates of NH₃-N and TP both reach more than 90 %, respectively. Moreover, the proposed technique is considerably effective in the removal of methanol.     

Bioremediation of a polycyclic aromatic hydrocarbon‐contaminated soil by using different aerobic batch bioreactor systems

The intrinsic depuration capability of a soil contaminated by polycyclic aromatic hydrocarbons (PAHs) originating from a contaminated industrial site was evaluated in this study by using different aerobic batch bioreactors: a slurry‐phase bioreactor, a blade‐agitated bioreactor, and a rotary vessel bioreactor. For each bioreactor, the disappearance of 14 target PAHs and of the total extractable organic matter was monitored. The three treatments exhibited rapid and extensive removal of the PAHs, which disappeared at different degradation rates according to their molecular weight and aromaticity degree. PAHs with two, three, and four aromatic rings were degraded in sequence, with average rates that generally decreased as the number of molecule rings increased. A slight increase in the bacterial biomass concentration and significant CO₂ production were also observed during the time course of the treatments. Among the three treatments, the slurry‐phase system provides the most effective and fastest removal of the PAHs and the organic extractable matter. However, the semisolid‐phase systems exhibited PAH depletion, capabilities higher than those reported in the literature for soils with similar particle size distribution in solid‐phase conditions.     

Organic matter transformations and kinetics during sewage sludge composting in a two-stage system

The use of different proportions of rape straw and grass as amendments in the composting of dewatered sewage sludge from a municipal wastewater treatment plant was tested in a two-stage system (first stage, an aerated bioreactor and second stage, a periodically turned windrow). The composition of feedstock affected the temperature and organic matter degradation in the bioreactor and the formation of humic substances, especially humic acids (HA), during compost maturation in the windrow. The total HA content (the sum of labile and stable HA) increased according to first-order kinetics, whereas labile HA content was constant and did not exceed 12% of total HA. Δlog K of 1.0–1.1 indicated that HA was of R-type, indicating a low degree of humification. Temperature during composting was the main factor affecting polymerization of fulvic acids to HA and confirmed the value of the degree of polymerization, which increased only when thermophilic conditions were obtained.     

Isolation of an aboriginal bacterial community capable of utilizing cyanide,thiocyanate, and ammonia from metallurgical plant wastewater

An aboriginal bacterial community capable of degrading cyanide (10 mg/l) and thiocyanate (2 g/l) and eliminating ammonia (120 mg/l) had been isolated from recycled water samples after blast-furnace gas purification of a metallurgical plant wastewater. It was shown that the optimal conditions for this bacterial community were as follows: temperature, 34°C; pH, 8.8–9.0; available organic matter concentration (glucose equivalent), 5 g/l; and dissolved O₂ concentration, 8–10 mg/l. This aboriginal community was formed by the bacteria belonging to the genus Pseudomonas.     

Feasibility study of a cyclic anoxic/aerobic two-stage MBR for treating ABS resin manufacturing wastewater

This study investigated the feasibility and the treatment efficiency of a cyclic anoxic/aerobic two-stage MBR for treating polymeric industrial wastewater. The anoxic/aerobic hybrid MBR was operated without sludge withdrawal except sampling during the study. The results showed that the highest COD organic loading rate of 8.7 kg COD/m³ day from bioreactor was obtained at phase 3. The system achieved 97% BOD₅ and 89% COD removal. It also revealed that 93% of COD removal was contributed by bioreactor at phase 3 and the similar results happened to phases 1 and 2. The highest TN and TKN removals for each phase were 60, 74, 80% and 61, 74, 81%, respectively and limited by nitritation step. SEM images of nascent and fouled membranes were offered to evaluate the cleaning method. The system was operated for 174 days, resulting in high degradation rate, flexibility towards influent fluctuations and limited sludge production.     

Comparison between the evaluation of bacterial regrowth capability in a turbidimeter and biodegradable dissolved organic carbon bioreactor measurements in water

In recent years, two different approaches to the study of biodegradable organic matter in distribution systems have been followed. The assimilable organic carbon (AOC) indicates the portion of the dissolved organic matter used by bacteria and converted to biomass, which is directly measured as total bacteria, active bacteria or colony-forming units and indirectly as ATP or increase in turbidity. In contrast, the biodegradable dissolved organic carbon (BDOC) is the portion of the dissolved organic carbon that can be mineralized by heterotrophic micro-organisms, and it is measured as the difference between the inflow and the outflow of a bioreactor. In this study, at different steps in a water treatment plant, the bacterial regrowth capability was determined by the AOC method that measures the maximum growth rate by using a computerized Monitek turbidimeter. The BDOC was determined using a plug flow bioreactor. Measurements of colony-forming units and total organic carbon (TOC) evolution in a turbidimeter and of colony-forming units at the inflow/outflow of the bioreactor were also performed, calculating at all sampling points the coefficient yield ( Y = cfu/ΔTOC) in both systems. The correlations between the results from the bioreactor and turbidimeter have been calculated ; a high correlation level was observed between BDOC values and all the other parameters, except for Y calculated from bacterial suspension measured in the turbidimeter.     

Enhanced removal of organic matter and ammoniacal-nitrogen in a column experiment of tidal flow constructed wetland system

A tidal flow constructed wetland system was investigated for the removal of organic matter and ammoniacal-nitrogen from diluted piggery wastewater. The results demonstrated that the operation of tidal flow enhanced the transfer of oxygen into wetland matrices. The supply of oxygen by the operation (473 gO2/m2d) matched the demand for wastewater treatment. The overall oxygen consumption rate in the system was considerably higher than the typical rate obtainable in conventional wetlands; most oxygen being used for the decomposition of organic matter. Compared with conventional systems, the tidal flow system demonstrated greater efficiency in the removal of organic matter. Significant nitrification did not take place, although 27-48% ammonia was removed from the wastewater. Immobilization by microbial cells and adsorption were the likely routes to remove ammonia under the specific experiment conditions. Percentage removals of BOD5, NH4-N and SS increased after effluent recirculation at a ratio of 1:1 was employed.     

An Integrated System for the Bioremediation of Wastewater Containing Xenobiotics and Toxcic Metals

An integrated system for the biotreatment of acidic wastewaters containing both toxic metals and organics is presented. It consists of two bioprocess stages (i) an anaerobic, SRB stage (containing alkaline‐tolerant s ulfate‐ r educing b acteria) that at pH 8 (chosen to acclimatize the bacteria in the biomedium) produces high concentrations of total sulfide ions (more than 400 mg/L) which are added to the wastewater to precipitate the heavy metals out at pH 2 as metal sulfides, and (ii) an aerobic, acidophilic stage containing heterotrophic bacteria (WJB3) that degrade organic xenobiotics. The anaerobic system was comprised of a 4‐L fluidized bed bioreactor with immobilized SRB, a mixing tank, and a precipitation tank. The effluent from the bioreactor with a high concentration of sulfide ions was fed into a mixing tank where model wastewaters containing toxic metals and phenol at pH 2 were also fed at increasing loading rates until free metal ions could be detected in the precipitation tank outlet. Then the effluent from the precipitation tank outlet was fed into a 2.5‐L aerobic bioreactor in which phenol was degraded. In this research, 100 % removal efficiencies were obtained with wastewaters containing more than 400 mg/L metal ions and 900 mg/L phenol at a 6‐h HRT of the mixing tank.     

Anaerobic treatment of sulphate-rich wastewaters

Until recently, biological treatment of sulphate-rich wastewater was rather unpopular because of the production of H₂S under anaerobic conditions. Gaseous and dissolved sulphides cause physical-chemical (corrosion, odour, increased effluent chemical oxygen demand) or biological (toxicity) constraints, which may lead to process failure. Anaerobic treatment of sulphate-rich wastewater can nevertheless be applied successfully provided a proper treatment strategy is selected. The strategies currently available are discussed in relation to the aim of the treatment: i) removal of organic matter, ii) removal of sulphate or iii) removal of both. Also a whole spectrum of new biotechnological applications (removal of organic chemical oxygen demand, sulphur, nitrogen and heavy metals), recently developed based on a better insight in sulphur transformations, are discussed.     

Reactive black-5 azo dye treatment in suspended and attach growth sequencing batch bioreactor using different co-substrates

Treatment of textile wastewater is a big challenge because of diverse chemical composition, high chemical strength and color of the wastewater. In the present study, treatment of wastewater containing reactive black-5 azo dye was studied in anaerobic sequencing batch bioreactor (SBBR) using mixed liquor suspended solids (MLSS) from suspended and attach growth bioreactors. MLSS at concentration of 1000 mg/L and reactive black-5 azo dye at 100 mg/L were used. A culture (10⁸–10⁹ CFU/ml) of pre-isolated bacterial strains (Psychrobacter alimentarius KS23 and Staphylococcus equorum KS26)) capable of degrading azo dyes in mineral salt medium was used to accelerate the treatment process in bioreactor. Different combinations of sludge, culture and dye were used for treatment using different co-substrates. About 85% COD removal was achieved by consortium (MLSS + KS23 + KS26) after 24 h in attach growth bioreactor. Similarly, 92% color removal was observed with consortium in attach growth bioreactor compared to 85% color removal in suspended bioreactor. Addition of bacterial culture (20%, v/v) to the bioreactor could enhance the rate of color removal. This study suggests that biotreatment of wastewater containing textile dyes can be achieved more efficiently in the attach growth bioreactor using yeast extract as a co-substrate and MLSS augmented with dye-degrading bacterial strains.     

Analysis of bacteria communities in an up-flow fixed-bed (UFB) bioreactor for treating sulfide in hydrocarbon wastewater

An up-flow fixed-bed (UFB) bioreactor with patented functional polyurethane foam (FPUF) carriers was used to treat sulfide in hydrocarbon wastewater. Community compositions of autotrophic and heterotrophic bacteria were analyzed by polymerase chain reaction–denaturing gradient gel electrophoresis (PCR–DGGE). DGGE results showed that a relatively stable bacterial community composed of heterotrophic and autotrophic bacteria formed in the bioreactor by the end of experiment, which ensured 92–100% sulfide removal efficiencies. Furthermore, autotrophic genera of Thiobacillus and Thiomonas, as well as those of the heterotrophic genus of Acinetobacter survived and exhibited high sulfide oxidation activity under all three operational conditions. Different special genera were also observed under each operational condition, such as the halophilic genus of Nesterenkonia. In addition, a new genus of sulfide oxidation bacteria was found in the bioreactor, which had the ability to synthesize cytoplasm from organic compounds. These genera have wide applications for the treatment of sulfide in hydrocarbon wastewater.     

Heavy metal fractionation and organic matter stabilization in sewage sludge treatment wetlands

The presence of heavy metals in sludge stabilized in a reed bed system may affect its use for agricultural purposes. However, the environmental impact of sludge depends on the availability and phytotoxicity of these heavy metals.The aim of this research was to determine the effectiveness of a reed bed (Phragmites australis) sludge treatment system in two urban wastewater treatment plants in Italy after a three-year period of operation: (i) by estimating the process of sludge stabilization, following conventional and nonconventional parameters related to the evolution of organic matter quality (water soluble carbon, dehydrogenase activity, pyrolytic fragments); (ii) by following the heavy metal bioavailability in the sludge through their fractionation. For heavy metal fractionation, the Community Bureau of Reference (BCR) was followed.The results showed that there was mineralization and stabilization of sludge over time, suggested by the decrease of about 35% in water soluble carbon and of about 60-80% of dehydrogenase activity. Moreover, significant values of benzene (17%), toluene (31%) and phenol (9%) were found at the end of experimentation in both treatment wetlands, highlighting the re-synthesis of humic-like matter.The results also showed that the content of heavy metals after 30 months was associated with the less mobile fractions of the sludge (more than 60% of total heavy metal content for almost metal), in particular, the fraction linked to the organic matter.     

Optimization and/or acclimatization of activated sludge process under heavy metals stress

The present study aimed to overcome the toxicity of the heavy metals load, discharged with the industrial effluents into Alexandria sewerage network, on the activated sludge treatment system through effective acclimation for organic matter and heavy metals removal. Optimization and/or acclimatization of the activated sludge process in the presence of Cu, Cd, Co and Cr contaminating mixed domestic-industrial wastewater was investigated. Acclimatization process was performed through abrupt and stepwise addition of tested metals using sequencing batch reactors treatment approach and evaluated as microbial oxygen uptake rate (OUR), dehydrogenase activity (DHA), organic matter (COD) and heavy metals removal. Abrupt addition of metals adversely affected sludge bioactivity leading to decline in the removal efficiency of the targeted contaminants and loss of floc structure. Metals IC₅₀ confirmed that copper possessed the highest toxicity towards the OUR, DHA activity and COD removal with orders Cu > Cd > Cr > Co; Cu > Cd > Co = Cr and Cu > Cd > Cr > Co, respectively. The highest metal removal was recorded for Cd followed by Co, Cu and finally Cr, most of which was retained in the dissolved influent. However, controlled stepwise application of the tested metals exhibited high sensitivity of DHA and OUR activities only at the highest metal concentrations although enhanced at the lowest concentrations while COD removal was not significantly affected. In conclusion, this approach resulted in adaptation of the system where sludge microbes acquired and developed natural resistance to such metals leading to remarkable enhancement of both organic matter and heavy metals removal.     

Metal-induced inhibition of anaerobic metabolism of volatile fatty acids and hydrogen

The effects of copper (Cu), chromium (Cr), cadmium (Cd), lead (Pb) and zinc (Zn) on the biotransformation of organic acids (acetate, propionate and butyrate) and H₂ were assessed in serum-bottle microcosms. Experiments were performed over a range of metal concentrations (20–200 mg/1) using biomass from an anaerobic bioreactor fed continuously with ethanol distillery waste as inoculum. In general, the added metals inhibited the biotransformation of organic acids with increasing metal concentration. However, the extent of inhibition varied for the different biotransformations and for the different metals tested. For example, the concentration of CuCl₂ effecting a 50% reduction in the rate constant for biotransformation of acetate, propionate and butyrate was 60, 75 and 30 mg/1, respectively. Cu and Cr (VI) were the most inhibitory metals in organic acid transformation, whereas Pb was the least toxic. The rate of biotransformation of acetate was reduced by half at Cu and Cr concentrations of 60 and 40 gm/1 respectively, whereas Cd, Pb, and Zn concentrations of 160 to 200 mg/l had little effect. The activities of hydrogenotrophic methanogens were much less affected by the same metals and metal concentrations.     

Isolation of an aboriginal bacterial community capable of utilizing cyanide, thiocyanate, and ammonia from metallurgical plant wastewater

An aboriginal bacterial community capable of degrading cyanide (10 mg/l) and thiocyanate (2 g/l) and eliminating ammonia (120 mg/l) had been isolated from recycled water samples after blast-furnace gas purification of a metallurgical plant wastewater. It was shown that the optimal conditions for this bacterial community were as follows: temperature, 34 degrees C; pH, 8.8-9.0; available organic matter concentration (glucose equivalent), 5 g/l; and dissolved O2 concentration, 8-10 mg/l. This aboriginal community was formed by the bacteria belonging to the genus Pseudomonas.     

Occurrence of Pathogenic Free‐Living Amoebae and Bacterial Indicators in a Constructed Wetland Treating Domestic Wastewater from a Single Household

Constructed wetlands are effective wastewater treatment systems because of their ability to remove large amounts of organic matter and pathogens. The goals of this study were to characterize the presence of pathogenic free‐living amoebae and bacterial indicators (total and fecal coliforms), and to ascertain the removal efficiencies of physical and chemical pollutants, in a constructed wetland treating domestic wastewater from a single household. Influent and effluent samples were collected monthly over a ten‐month period for biological, physical and chemical analyses. Thirty‐two species of free‐living amoebae were isolated from the system. The genus Acanthamoeba was the most frequently encountered (59 %) and was removed from the wastewater with the greatest efficiency (80 %). Removal of bacteria was low, the highest removal rates were found in August (4 logarithmic units) and January (3 logarithmic units). The average removal efficiencies of suspended solids, BOD₅ and ammoniacal nitrogen were 71.5 %, 50.6 % and 13.1 %, respectively. The relatively low removal efficiencies of the various bacteriological, physical and chemical parameters suggest that the hydraulic retention time was probably insufficient for optimal treatment to occur. The effluent quality was unacceptable for unrestricted irrigation of crops that are eaten uncooked.     

Hectare-scale demonstration of high rate algal ponds for enhanced wastewater treatment and biofuel production

High rate algal ponds (HRAPs) are shallow, paddlewheel-mixed open raceway ponds that are an efficient and cost-effective upgrade for the conventional wastewater treatment ponds used by communities and farms the world over. HRAPs provide improved natural disinfection and nutrient removal and can be further enhanced by carbon dioxide (CO₂) addition to promote algal growth which is often carbon limited. This paper discusses the construction and operation of a 5-ha demonstration HRAP system treating primary settled wastewater at the Christchurch wastewater treatment plant, New Zealand. The system consisted of four 1.25-ha HRAPs that were constructed from an existing conventional pond. Algae were harvested from the HRAP effluent in specially designed settlers, which concentrated the algal/bacterial biomass to 1–2% organic solids for conversion to bio-crude oil following dewatering. Performance data from the first 15?months of HRAP operation (without CO₂ addition) are presented. The four demonstration HRAPs had reasonable replication of both treatment performance and algal/bacterial productivity with similar annual average wastewater treatment efficiency (~50% removal of BOD₅, ~87% removal of fBOD₅, ~65% removal of ammoniacal-N, ~19% removal of dissolved reactive phosphorus and ~2 log removal of Escherichia coli), algal species composition and algal/bacterial biomass production (~8?g?m^?2?day ?1 volatile suspended solids). These results were in good agreement with the results for pilot-scale HRAP without CO₂ addition in New Zealand. This study provides further indication of the potential for energy efficient and effective wastewater treatment using HRAP, while biofuel conversion of the harvested algal bacterial biomass could provide a valuable niche distributed energy source for local communities.