Bioaugmentation of cyanide-degrading microorganisms in a full-scale cokes wastewater treatment facility

To enhance biological removal efficiency of total cyanides, bioaugmentation was applied to a full-scale cokes wastewaters treatment process. After a laboratorial-scale cultivation (up to 1.2 m(3)) of a cyanide-degrading yeast (Cryptococcus humicolus) and unidentified cyanide-degrading microorganisms, the microbial consortium was inoculated into a fluidized-bed type process (1280 m(3)), and then enriched for two months with a huge supply of glucose, KCN and other nutrients. Target wastewater was effluent of a biological pre-denitrification process for treating cokes wastewater, and contained about 14 mg/L of total cyanides in the form of ferric cyanide. This may be a first or rare report on the full-scale bioaugmentation of specialized-microorganisms. However, continuous operation of the full-scale cyanides-degrading bioprocess showed poor removal efficiency than expected owing to poor settling performance of microbial flocs, slow biodegradation rate of ferric cyanide and lack of organic carbon sources within the wastewater. Therefore, there is a need for further studies on how to solve these operating problems in full-scale bioaugmentation approach.     

Calibration of a complex activated sludge model for the full-scale wastewater treatment plant

In this study, the results of the calibration of the complex activated sludge model implemented in BioWin software for the full-scale wastewater treatment plant are presented. Within the calibration of the model, sensitivity analysis of its parameters and the fractions of carbonaceous substrate were performed. In the steady-state and dynamic calibrations, a successful agreement between the measured and simulated values of the output variables was achieved. Sensitivity analysis revealed that upon the calculations of normalized sensitivity coefficient (S _i,j ) 17 (steady-state) or 19 (dynamic conditions) kinetic and stoichiometric parameters are sensitive. Most of them are associated with growth and decay of ordinary heterotrophic organisms and phosphorus accumulating organisms. The rankings of ten most sensitive parameters established on the basis of the calculations of the mean square sensitivity measure (δ _j ^msqr) indicate that irrespective of the fact, whether the steady-state or dynamic calibration was performed, there is an agreement in the sensitivity of parameters.     

Sequential modelling of a full-scale wastewater treatment plant using an artificial neural network

This work proposes a sequential modelling approach using an artificial neural network (ANN) to develop four independent multivariate models that are able to predict the dynamics of biochemical oxygen demand (BOD), chemical oxygen demand (COD), suspended solid (SS), and total nitrogen (TN) removal in a wastewater treatment plant (WWTP). Suitable structures of ANN models were automatically and conveniently optimized by a genetic algorithm rather than the conventional trial and error method. The sequential modelling approach, which is composed of two parts, a process disturbance estimator and a process behaviour predictor, was also presented to develop multivariate dynamic models. In particular, the process disturbance estimator was first employed to estimate the influent quality. The process behaviour predictor then sequentially predicted the effluent quality based on the estimated influent quality from the process disturbance estimator with other process variables. The efficiencies of the developed ANN models with a sequential modelling approach were demonstrated with a practical application using a data set collected from a full-scale WWTP during 2 years. The results show that the ANN with the sequential modelling approach successfully developed multivariate dynamic models of BOD, COD, SS, and TN removal with satisfactory estimation and prediction capability. Thus, the proposed method could be used as a powerful tool for the prediction of complex and nonlinear WWTP performance.     

Methane emissions from a full-scale A/A/O wastewater treatment plant

Methane (CH₄) emissions from a full-scale anaerobic/anoxic/oxic (A/A/O) wastewater treatment plant (WWTP) (Jinan, China) were investigated during spring and summer of 2010. Results showed that the major emission sources of CH₄ performed the following descending order: anaerobic tanks, oxic tanks, aerated grit chambers and sludge concentration tanks. The total annual fluxes of CH₄ emissions from the Jinan WWTP were 1.69 × 10⁴ kg yr⁻¹, with the emission factors of per capita emissions of 11.3 g CH₄ person⁻¹ yr⁻¹ and flow-based emissions of 1.55 × 10⁻⁴ g CH₄ (L of wastewater)⁻¹. The estimated source strength of methane for all WWTPs in China was 6.2 Gg yr⁻¹ (1 Gg = 10⁹ g). The most significant factors influencing methane emissions were dissolved oxygen concentration in aerated grit chamber and oxic tank and water temperature in high density settler tanks.     

Microbial biogeography across a full-scale wastewater treatment plant transect: evidence for immigration between coupled processes

Wastewater treatment plants use a variety of bioreactor types and configurations to remove organic matter and nutrients. Little is known regarding the effects of different configurations and within-plant immigration on microbial community dynamics. Previously, we found that the structure of ammonia-oxidizing bacterial (AOB) communities in a full-scale dispersed growth activated sludge bioreactor correlated strongly with levels of NO₂ ^? entering the reactor from an upstream trickling filter. Here, to further examine this puzzling association, we profile within-plant microbial biogeography (spatial variation) and test the hypothesis that substantial microbial immigration occurs along a transect (raw influent, trickling filter biofilm, trickling filter effluent, and activated sludge) at the same full-scale wastewater treatment plant. AOB amoA gene abundance increased >30-fold between influent and trickling filter effluent concomitant with NO₂ ^? production, indicating unexpected growth and activity of AOB within the trickling filter. Nitrosomonas europaea was the dominant AOB phylotype in trickling filter biofilm and effluent, while a distinct “Nitrosomonas-like” lineage dominated in activated sludge. Prior time series indicated that this “Nitrosomonas-like” lineage was dominant when NO₂ ^? levels in the trickling filter effluent (i.e., activated sludge influent) were low, while N. europaea became dominant in the activated sludge when NO₂ ^? levels were high. This is consistent with the hypothesis that NO₂ ^? production may cooccur with biofilm sloughing, releasing N. europaea from the trickling filter into the activated sludge bioreactor. Phylogenetic microarray (PhyloChip) analyses revealed significant spatial variation in taxonomic diversity, including a large excess of methanogens in the trickling filter relative to activated sludge and attenuation of Enterobacteriaceae across the transect, and demonstrated transport of a highly diverse microbial community via the trickling filter effluent to the activated sludge bioreactor. Our results provide compelling evidence that substantial immigration between coupled process units occurs and may exert significant influence over microbial community dynamics within staged bioreactors.     

Impact of operational parameters on bacterial community in a full-scale municipal wastewater treatment plant

A bacterial community in activated sludge from a full-scale municipal wastewater treatment plant was monitored throughout the year with the use of FISH, RISA and DGGE techniques. In the investigated range of temperatures (11.9-21.6 degrees C), a rise in temperature resulted in a lower total bacteria richness, while organic load rate changes from 0.09 to 0.21 g COD x g TSS(-1) x d(-1) were positively correlated with the number of bands in RISA patterns. The most diverse pattern (29 different bands) was characteristic for the activated sludge sample collected at the end of January at wastewater temperature of 11.9 degrees C. The ammonia-oxidising bacteria community did not change during the study, and comprised of 4 different bacterial populations with one dominant species closely related to Nitrosospira sp. REGAU (GenBank accession number AY635572.1). The percentage of ammonia-oxidising bacteria in the activated sludge varied from 6.2 to 19.5% and depended on temperature (R = 0.61, p = 0:02) and organic load rate (R = -0.55, p = 0.04).     

Emergence of competitive dominant ammonia-oxidizing bacterial populations in a full-scale industrial wastewater treatment plant

Ammonia-oxidizing bacterial populations in an industrial wastewater treatment plant were investigated with amoA and 16S rRNA gene real-time PCR assays. Nitrosomonas nitrosa initially dominated, but over time RI-27-type ammonia oxidizers, also within the Nitrosomonas communis lineage, increased from below detection to codominance. This shift occurred even though nitrification remained constant.     

Dynamics of the microfauna community in a full-scale municipal wastewater treatment plant experiencing sludge bulking

We investigated the dynamics of the microfauna community in activated sludge, with special reference to sludge bulking, in two parallel municipal wastewater treatment systems in Beijing, China over a period of 14 months. Annual cyclic changes in microfauna community structures occurred in both systems. RELATE analysis based on Spearman's Rank correlation indicated that microfauna community structures were highly correlated with the sludge volume index (SVI) (p < 0.001), which indicates sludge settleability. Nutrient conditions of raw sewage (p < 0.01) and hydraulic retention time (HRT) (p < 0.05) were also related to microfauna community structures. Abundances of the species Epistylis plicatilis and Vorticella striata increased significantly with an increase in SVI (p < 0.001) and decrease in water temperature (p < 0.001), suggesting that sludge bulking may have created favorable conditions for the two species, even under unfavorable temperature conditions. Sludge de-flocculation primarily due to the excessive growth of Microthrix parvicella-like filaments could be an important driving force for the microfauna community changes. The release of flocculated non-filamentous bacteria may represent a suitable food source for these species. The two species may be considered as potential bioindicators for sludge bulking.     

Bacterial communities in full-scale wastewater treatment systems

Bacterial metabolism determines the effectiveness of biological treatment of wastewater. Therefore, it is important to define the relations between the species structure and the performance of full-scale installations. Although there is much laboratory data on microbial consortia, our understanding of dependencies between the microbial structure and operational parameters of full-scale wastewater treatment plants (WWTP) is limited. This mini-review presents the types of microbial consortia in WWTP. Information is given on extracellular polymeric substances production as factor that is key for formation of spatial structures of microorganisms. Additionally, we discuss data on microbial groups including nitrifiers, denitrifiers, Anammox bacteria, and phosphate- and glycogen-accumulating bacteria in full-scale aerobic systems that was obtained with the use of molecular techniques, including high-throughput sequencing, to shed light on dependencies between the microbial ecology of biomass and the overall efficiency and functional stability of wastewater treatment systems. Sludge bulking in WWTPs is addressed, as well as the microbial composition of consortia involved in antibiotic and micropollutant removal.     

Sudden failure of biological nitrogen and carbon removal in the full-scale pre-denitrification process treating cokes wastewater

A full-scale pre-denitrification process treating cokes wastewater containing toxic compounds such as phenols, cyanides and thiocyanate has shown good performance in carbon and nitrogen removal. However, field operators have been having trouble with its instability without being able to identify the causes. To clarify the main cause of these sudden failures of the process, comprehensive studies were conducted on the pre-denitrification process using a lab-scale reactor system with real cokes wastewater. First, the shock loading effects of three major pollutants were investigated individually. As the loading amount of phenol increased to 600 mg/L, more COD, TOC and phenol itself were flowed into the aerobic reactor, but phenol itself did not inhibit nitrification and denitrification, owing to the effect of dilution and its rapid biodegradation. Higher loading of ammonia or thiocyanate slightly enhanced the removal efficiency of organic matter, but caused the final discharge concentration of total nitrogen to be above its legal limit of 60 mg-N/L. Meanwhile, continuous inflow of abnormal wastewater collected during unstable operation of the full-scale pre-denitrification process, caused a sudden failure of nitrogen removal in the lab-scale process, like the removal pattern of the full-scale one. This was discovered to be due to the lack of inorganic carbon in the aerobic reactor where autotrophic nitrification occurs.     

Performance and spatial succession of a full-scale anaerobic plant treating high-concentration cassava bioethanol wastewater

A novel two-phase anaerobic treatment technology was developed to treat high-concentration organic cassava bioethanol wastewater. The start-up process and contribution of organics (COD, total nitrogen, and NH4 +-N) removal in spatial succession of the whole process and spatial microbial diversity changing when sampling were analyzed. The results of the start-up phase showed that the organic loading rate could reach up to 10 kg COD/m(3)d, with the COD removal rate remaining over 90% after 25 days. The sample results indicated that the contribution of COD removal in the pre-anaerobic and anaerobic phases was 40% and 60%, respectively, with the highest efficiency of 98.5%; TN and NH4 +-N had decreased to 0.05 g/l and 0.90 g/l, respectively, and the mineralization rate of total nitrogen was 94.8%, 76.56% of which was attributed to the anaerobic part. The microbial diversity changed remarkably among different sample points depending on the physiological characteristics of identified strains. Moraxellaceae, Planococcaceae, and Prevotellaceae were dominant in the pre-anaerobic phase and Bacteroidetes, Campylobacterales, Acinetobacter, Lactobacillus, Clostridium, and Bacillus for the anaerobic phase. Methanosarcinaceae and Methanosaeta were the two main phylotypes in the anaerobic reactor.     

R-plasmid transfer in a wastewater treatment plant.

Enteric bacteria have been examined for their ability to transfer antibiotic resistance in a wastewater treatment plant. Resistant Salmonella enteritidis, Proteus mirabilis, and Escherichia coli were isolated from clinical specimens and primary sewage effluent. Resistance to ampicillin, chloramphenicol, streptomycin, sulfadiazine, and tetracycline was demonstrated by spread plate and tube dilution techniques. Plasmid mediation of resistance was shown by ethidium bromide curing, agarose gel electrophoresis, and direct cell transfer. Each donor was mated with susceptible E. coli and Shigella sonnei. Mating pairs (and recipient controls) were suspended in unchlorinated primary effluent that had been filtered and autoclaved. Suspensions were added to membrane diffusion chambers which were then placed in the primary and secondary setting tanks of the wastewater treatment plant. Resistant recombinants were detected by replica plating nutrient agar master plates onto xylose lysine desoxycholate agar plates that contained per milliliter of medium 10 micrograms of ampicillin, 30 micrograms of chloramphenicol, 10 micrograms of streptomycin, 100 micrograms of sulfadiazine, or 30 micrograms of tetracycline. Mean transfer frequencies for laboratory matings were 2.1 X 10(-3). In situ matings for primary and secondary settling resulted in frequencies of 4.9 X 10(-5) and 7.5 X 10(-5), respectively. These values suggest that a significant level of resistance transfer occurs in wastewater treatment plants in the absence of antibiotics as selective agents.     

Environmental performance of a municipal wastewater treatment plant

Goal, Scope and Background  Nowadays, every strategy must be developed taking into account the global impact on the environment; if this aspect is forgotten, a change of environmental loads or their effect will be caused and no reduction will be attained. For instance, a wastewater treatment plant (WWIP), which is considereda priori as an ecological treatment system, gives rise to an environmental impact due to its energy consumption, use of chemical compounds, emissions to the atmosphere and sludge production, the post-treatment of which will also have diverse environmental effects. The goal of this study is to evaluate the potential environmental impact corresponding to a municipal WW1P and to identify the hot spots associated with the process. Methods  In this study, the Centre of Environmental Science (CML) of Leiden University methodology has been considered to quantify the potential environmental impact associated with the system under study. A comprehensive analysis of the WWTP was evaluated for the physico-chemical characterisation of the wastewaters as well as the inventory of all the inputs (energy, chemical compounds, ...) and outputs (emissions to air, water, soil and solid waste generation) associated with the global process. Regarding Life Cycle Inventory Assessment, SimaPro 5.0 was used and in particular CML factors (updated in 2002) were chosen for characterisation and normalisation stages. Results and Discussion  A comprehensive inventory of empirical data from water, sludge and gas flows during 2000 and 2001 was obtained. Two impact categories arise due to their significance: eutrophication and terrestrial ecotoxicity. Consequently, the aspects to be minimised in order to reduce the environmental impact of the system are the pollutant load at the watercourse discharge (mainly NH₃, PO₄ ^[3- and COD, even when all of them are below legal limits) and the emissions to soil (mainly Cr, Hg and Zn, even when they are present in low concentrations) when the sludge is used for agricultural application. Conclusions  As far as the environmental impact is concerned, differentiation between humid and dry season is not required as results are practically equal for both situations. Water discharge and sludge application to land have turned out to be the main contributors in the environmental performance of a WWTP. Regarding the former, the removal of nitrogen by means of a nitrification-denitrification system coupled to conventional biological aerobic treatment implies a high environmental impact reduction and, as for the latter, bearing in mind the proposed legislation, heavy metals as well as pathogens are supposed to be the key parameters to define the most adequate treatment strategies for the generated sludge. Recommendations and Outlook  This study can serve as a basis for future studies that can apply a similar policy to a great number of wastewater facilities. Besides, features such as different treatment systems and capacities can provide additional information with the final aim of including the environmental vector in the decision-making process when the operation of a WWTP is intended to be optimised. Moreover, sludge must also be a focus of attention due to the expected increase and its major contribution to the global environmental impact of a WWTP, which can determine other treatment alternatives.     

Enhancing nitrogen removal in an Orbal oxidation ditch by optimization of oxygen supply: practice in a full-scale municipal wastewater treatment plant

Seven different aeration modes, in which oxygen supply was changed by adjusting the number of aerators, were designed and applied in a full-scale municipal wastewater treatment plant with Orbal oxidation ditch to investigate the influence of dissolved oxygen (DO) on nitrogen removal performance. The full-scale experiment results of 574 days showed that nitrogen removal efficiency depended on the degree of nitrification and denitrification in the outer channel, which was the largest contributor for TN removal in the Orbal oxidation ditch. Appropriate aeration control in the outer channel was essential to balance nitrification and denitrification in the Orbal oxidation ditch. When DO was as low as about 0.2 mg/L in the outer channel, the highest TN removal efficiency of 75% was obtained. Microbial analysis confirmed that aerobic and anaerobic bacteria coexisted in the outer channel. The greater species diversity and more intensive activities of these bacteria in aeration Mode V may be responsible for the higher TN removal efficiency compared with Mode III. These results suggest that different aerated conditions in the Orbal oxidation ditch might have a significant effect on microbial community characteristics and nitrogen removal efficiencies.     

Ammonia-oxidizing archaea and ammonia-oxidizing bacteria in six full-scale wastewater treatment bioreactors

In this study, dideoxy sequencing and 454 high-throughput sequencing were used to analyze diversities of the ammonia monooxygenase (amoA) genes and the 16S rRNA genes of ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) in six municipal wastewater treatment plants. The results showed that AOB amoA genes were quite diverse in different wastewater treatment plants while the 16S rRNA genes were relatively conserved. Based on the observed complexity of amoA and 16S rRNA genes, most of the AOB can be assigned to the Nitrosomonas genus, with Nitrosomonas ureae, Nitrosomonas oligotropha, Nitrosomonas marina, and Nitrosomonas aestuarii being the four most dominant species. From the sequences of the AOA amoA genes, most AOA observed in this study belong to the CGI.1b group, i.e., the soil lineage. The AOB amoA and 16S rRNA genes were quantified by quantitative PCR and 454 high-throughput pyrosequencing, respectively. Although the results from the two approaches show some disconcordance, they both indicated that the abundance of AOB in activated sludge was very low.     

Molecular evidence for novel planctomycete diversity in a municipal wastewater treatment plant

We examined anoxic and aerobic basins and an anaerobic digestor of a municipal wastewater treatment plant for the presence of novel planctomycete-like diversity. Three 16S rRNA gene libraries were constructed by using a 16S rRNA-targeted universal reverse primer and a forward PCR primer specific for Planctomyces: Phylogenetic analysis of 234 16S rRNA gene sequences defined 110 operational taxonomic units. The majority of these sequences clustered with the four known genera, Pirellula (32%), Planctomyces (18.4%), Gemmata (3.8%), and Isosphaera (0.4%). More interestingly, 42.3% of the sequences appeared to define two distantly separated monophyletic groups. The first group, represented by 35.5% of the sequences, was related to the Planctomyces group and branched as a monophyletic cluster. It exhibited between 11.9 and 20.3% 16S rRNA gene sequence dissimilarity in comparisons with cultivated planctomycetes. The second group, represented by 6.8% of the sequences, was deeply rooted within the Planctomycetales tree. It was distantly related to the anammox sequences (level of dissimilarity, 20.3 to 24.4%) and was a monophyletic cluster. The retrieved sequences extended the intralineage phylogenetic depth of the Plantomycetales from 23 to 30.6%. The lineages described here may have a broad diversity of undiscovered biochemical and metabolic novelty. We developed a new 16S rRNA-targeted oligonucleotide probe and localized members of one of the phylogenetic groups using the fluorescent in situ hybridization technique. Our results indicate that activated sludge contains very diverse representatives of this group, which grow under aerobic and anoxic conditions and even under anaerobic conditions. The majority of species in this group remain poorly characterized.     

Involvement of Rhodocyclus-related organisms in phosphorus removal in full-scale wastewater treatment plants

The participation of organisms related to Rhodocyclus in full-scale enhanced biological phosphorus removal (EBPR) was investigated. By using fluorescent in situ hybridization techniques, the communities of Rhodocyclus-related organisms in two full-scale wastewater treatment plants were estimated to represent between 13 and 18% of the total bacterial population. However, the fractions of these communities that participated in polyphosphate accumulation depended on the type of treatment process evaluated. In a University of Cape Town EBPR process, the percentage of Rhodocyclus-related cells that contained polyphosphate was about 20% of the total bacterial population, but these cells represented as much as 73% of the polyphosphate-accumulating organisms (PAOs). In an aerated-anoxic EBPR process, Rhodocyclus-related PAOs were less numerous, accounting for 6% of the total bacterial population and 26% of the total PAO population. In addition, 16S ribosomal DNA sequences 99.9% similar to the sequences of Rhodocyclus-related organisms enriched in acetate-fed bench-scale EBPR reactors were recovered from both full-scale plants. These results confirmed the involvement of Rhodocyclus-related organisms in EBPR and demonstrated their importance in full-scale processes. In addition, the results revealed a significant correlation between the type of EBPR process and the PAO community.     

Reduction in excess sludge production in a dairy wastewater treatment plant via nozzle-cavitation treatment: Case study of an on-farm wastewater treatment plant

Nozzle-cavitation treatment was used to reduce excess sludge production in a dairy wastewater treatment plant. During the 450-d pilot-scale membrane bioreactor (MBR) operation, when 300 l of the sludge mixed liquor (1/10 of the MBR volume) was disintegrated per day by the nozzle-cavitation treatment with the addition of sodium hydrate (final concentration: 0.01% W/W) and returned to the MBR, the amount of excess sludge produced was reduced by 80% compared with that when sludge was not disintegrated.