A comparative study of gel entrapped and membrane attached microbial reactors for biodegrading phenol

A comparative study between two reactors, one using microorganisms entrapped in calcium alginate gel, and the other using microorganisms attached on the surface of a membrane (polymeric microporous sheeting, MPS^TM) to biodegrade phenol is performed. Results indicate that the alginate bead bioreactor is efficient at higher phenol concentrations while the membrane bioreactor shows better performance at lower phenol concentrations. This unique response is primarily attributed to the different techniques by which the microorganisms are immobilized in the two reactors.In batch mode, below a starting concentration of 100 ppm phenol, biodegradation rates in the membrane bioreactor are (7.58 to 12.02 mg phenol/h · g dry biomass) atleast 10 times the rates in alginate bead bioreactor (0.74 to 1.32 mg phenol/h · g dry biomass). Biodegradation rates for the two reactors match at a starting concentration of 250 ppm phenol. Above 500 ppm phenol, the rates in the alginate bead bioreactor are (7.3 to 8.1 mg phenol/h · g dry biomass) on an average 5.5 times the corresponding rates in the membrane bioreactor (2.18 to 1.03 mg phenol/h · g dry biomass).In continuous feed mode the steady state degradation rates in the membrane bioreactor are one to two orders of magnitude higher than the alginate bead bioreactor below 150 ppm inlet phenol concentration. At an inlet concentration around 250 ppm phenol the rates are comparable. Above 500 ppm of phenol the rates in the alginate bioreactor are an order of magnitude high than the membrane bioreactor.Due to substrate inhibition, and its inability to sustain a high biomass concentration, the membrane bioreactor shows poor efficiencies at phenol concentrations above 250 ppm. At low phenol concentrations the apparent reaction rates in the alginate bead bioreactor decrease due to the diffusional resistance of the gel matrix, while biodegradation rates in the membrane bioreactor remain high due to essentially no external diffusional resistance.Results indicate that a combined reactor system can be more effective for bioremediation than either separate or attached microbial reactors.     

Aggregation of immobilized activated sludge cells into aerobically grown microbial granules for the aerobic biodegradation of phenol

AIMS: The aim of this study is to evaluate the utility of aerobically grown microbial granules for the biological treatment of phenol-containing wastewater. METHODS AND RESULTS: A column-type sequential aerobic sludge blanket reactor was inoculated with activated sludge and fed with phenol as the sole carbon source, at a rate of 1.5 g phenol l-1 d-1. Aerobically grown microbial granules first appeared on day 9 of reactor operation and quickly grew to displace the seed flocs as the dominant form of biomass in the reactor. These granules were compact and regular in appearance, and consisted of bacterial rods and cocci and fungi embedded in an extracellular polymeric matrix. The granules had a mean size of 0.52 mm, a sludge volume index of 40 ml g-1 and a specific oxygen utilization rate of 110 mg oxygen g VSS-1 h-1 (VSS stands for volatile suspended solids). Specific phenol degradation rates increased with phenol concentration from 0 to 500 mg phenol l-1, peaked at 1.4 g phenol g VSS-1 d-1, and declined with further increases in phenol concentration as substrate inhibition effects became important. CONCLUSIONS: Aerobically grown microbial granules were successfully cultivated in a reactor maintained at a loading rate of 1.5 g phenol l-1 d-1. The granules exhibited a high tolerance towards phenol. Significant rates of phenol degradation were attained at phenol concentrations as high as 2 g l-1. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study to demonstrate the ability of aerobically grown microbial granules to degrade phenol. These granules appear to represent an excellent immobilization strategy for microorganisms to biologically remove phenol and other toxic chemicals in high-strength industrial wastewaters.     

Scale-up impacts on mass transfer and bioremediation of suspended naphthalene particles in bead mill bioreactors

Scale-up effects on mass transfer and bioremediation of suspended naphthalene particles have been studied in 20 and 58L bead mill bioreactors and compared to data generated earlier with a laboratory scaled bioreactor. The bead mill bioreactor performance with respect to naphthalene mass transfer rate was dependent on the size and loading of the inert particles, as well as the rotational speed of the roller apparatus. The optimum operating conditions were found to be 15mm glass beads at a loading of 50% (total volume of particles/working volume of bioreactor: v/v%) and a bioreactor rotational speed of 50rpm. The highest naphthalene mass transfer coefficients obtained in the large scale system under these optimum conditions (19.6 and 22.4h(-1) for 20 and 58L vessels, respectively) were higher than those determined previously in a 2.5L bead mill bioreactor (0.7h(-1)). The acute toxicity tests indicated that the bioreactor effluent was less toxic than the untreated naphthalene suspension. Biodegradation rates obtained in these large scale bead mill bioreactors under optimum conditions (36-37.4mgL(-1)h(-1)) were higher than those achieved in the control bioreactors of similar sizes (11.4 and 11.6mgL(-1)h(-1)) but were slower than those previously determined in a 2.5L bead mill bioreactor (59-61.5mgL(-1)h(-1)). The limitation of oxygen in the large scale systems and damage of the bacterial cells due to the crushing effects of the large beads are likely contributing factors in the lower observed biodegradation rates. The optimum conditions with respect to naphthalene mass transfer might not necessarily translate to optimum performance with regard to bioremediation.     

Feasibility studies on the treatment of dairy wastewaters with upflow anaerobic sludge blanket reactors

The feasibility of using upflow anaerobic sludge blanket (UASB) reactors for the treatment of dairy wastewaters was explored. Two types of UASBs were used--one operating on anaerobic sludge granules developed by us from digested cowdung slurry (DCDS) and the other on the granules obtained from the reactors of M/s EID Parry treating sugar industry wastewaters. The reactors were operated at HRT of 3 and 12 h and on COD loading rates ranging from 2.4 kg per m3 of digester volume, per day to 13.5 kg m(-3) d(-1). At the 3 h HRT, the maximum COD reduction in the DCDS-seeded and the industrial sludge-seeded reactors was 95.6% and 96.3%, respectively, better than at 12 h HRT (90% and 92%, respectively). In both the reactors, the maximum, the second best, and the third best COD reduction occurred at the loading rates of 10.8, 8.6 and 7.2 kg m3 d(-1), respectively. At loading rates higher than 10.8 kg, the reactor performance dropped precipitously. Whereas in the first few months the reactors operating on sludge from EID Parry achieved better biodegradation of the waste, compared to the reactors operated on DCDS, the performance of the latter gradually improved and matched with the performance of the former.     

Start-up of the Anammox process in a membrane bioreactor

The start-up of an Anammox process was studied in a membrane sequencing batch reactor (MSBR) in which a submerged hollow fibre membrane module was used to retain the biomass. The reactor was seed with Anammox biomass and fed using the Van de Graaf medium. During a first operating stage, salt precipitation was observed and interfered with microbial activity and caused a decrease of the nitrogen removal rate of the reactor from 100 to only 10 mgl(-1) per day. Salt precipitation was avoided by diminishing adequately the Ca and P concentrations of the Van de Graaf medium during the last operating stage. This action increased quickly the activity of the system, and nitrogen removal rate reached up to 710 mgl(-1) per day with almost full nitrite removal. Sporadic flotation of the sludge was observed in the MSBR. The use of the membrane avoided biomass wash-out from the system. Moreover, a surprising fact was that Anammox biomass did not grow in flocs in the MSBR, but in granules. This fact showed that this kind of microorganisms have a trend to grow in aggregates. Results indicated that the use of the MSBR could be a suitable system for nitrogen removal by using the Anammox reaction.     

Biodegradation of methyl tert-butyl ether as a sole carbon source by aerobic granules cultivated in a sequencing batch reactor

Aerobic granules efficient at degrading methyl tert-butyl ether (MTBE) were successfully developed in a well-mixed sequencing batch reactor (SBR). Treatment efficiency of MTBE in the reactor during the stable operations exceeded 99.8%, and effluent MTBE was consistently below 800 mug/L. The specific MTBE degradation rate was observed to increase with increasing MTBE initial concentrations from 25 to 400 mg/L, peaked at 18.2 mg-MTBE/g-VSS h, and declined with further increases in MTBE concentration as substrate inhibition effects became significant. There was a good fit between these biodegradation data and the Haldane equation (R (2) = 0.976). Microbial community DNA profiling was carried out using denaturing gradient gel electrophoresis (DGGE) of polymerase chain reaction amplified 16S rDNA. The aerobic granule was found to contain a wide diversity of microorganisms. More than 70% similarity among the samples in the time period examined indicated a highly stable microbial community as the reactor reached the stable operation.     

Empirical model for the autotrophic biodegradation of thiocyanate in an activated sludge reactor

AIMS: The aim of this investigation was to develop an empirical model for the autotrophic biodegradation of thiocyanate using an activated sludge reactor. METHODS AND RESULTS: The methods used for this purpose included the use of a laboratory scale activated sludge reactor unit using thiocyante feed concentrations from 200 to 550 mg x l(-1). Reactor effluent concentrations of <1 mg x l(-1) thiocyanate were consistently achieved for the entire duration of the investigation at a hydraulic retention time of 8 h, solids (biomass) retention of 18 h and biomass (dry weight) concentrations ranging from 2 to 4 g x l(-1). A biomass specific degradation rate factor was used to relate thiocyanate degradation in the reactor to the prevailing biomass and thiocyanate feed concentrations. A maximum biomass specific degradation rate of 16 mg(-1) x g(-1) x h(-1) (mg thiocyanate consumed per gram biomass per hour) was achieved at a thiocyanate feed concentration of 550 mg x l(-1). The overall yield coefficient was found to be 0.086 (biomass dry weight produced per mass of thiocyanate consumed). CONCLUSION: Using the results generated by this investigation, an empirical model was developed, based on thiocyanate feed concentration and reactor biomass concentration, to calculate the required absolute hydraulic retention time at which a single-stage continuously stirred tank activated sludge reactor could be operated in order to achieve an effluent concentration of <1 mg x l(-1). The use of an empirical model rather than a mechanistic-based kinetic model was proposed due to the low prevailing thiocyanate concentrations in the reactor. SIGNIFICANCE AND IMPACT OF THE STUDY: These results represent the first empirical model, based on a comprehensive data set, that could be used for the design of thiocyanate-degrading activated sludge systems.     

Biodegradation and kinetics of aerobic granules under high organic loading rates in sequencing batch reactor

Biodegradation, kinetics, and microbial diversity of aerobic granules were investigated under a high range of organic loading rate 6.0 to 12.0 kg chemical oxygen demand (COD) m⁻³ day⁻¹ in a sequencing batch reactor. The selection and enriching of different bacterial species under different organic loading rates had an important effect on the characteristics and performance of the mature aerobic granules and caused the difference on granular biodegradation and kinetic behaviors. Good granular characteristics and performance were presented at steady state under various organic loading rates. Larger and denser aerobic granules were developed and stabilized at relatively higher organic loading rates with decreased bioactivity in terms of specific oxygen utilization rate and specific growth rate (μ _overall) or solid retention time. The decrease of bioactivity was helpful to maintain granule stability under high organic loading rates and improve reactor operation. The corresponding biokinetic coefficients of endogenous decay rate (k _d), observed yield (Y _obs), and theoretical yield (Y) were measured and calculated in this study. As the increase of organic loading rate, a decreased net sludge production (Y _obs) is associated with an increased solid retention time, while k _d and Y changed insignificantly and can be regarded as constants under different organic loading rates.     

Bioremediation of gasoline-contaminated groundwater in a pilot-scale packed-bed anaerobic reactor

This work reports on the anaerobic treatment of gasoline-contaminated groundwater in a pilot-scale horizontal-flow anaerobic immobilized biomass reactor inoculated with a methanogenic consortium. BTEX removal rates varied from 59 to 80%, with a COD removal efficiency of 95% during the 70 days of in situ trial. BTEX removal was presumably carried out by microbial syntrophic interactions, and at the observed concentrations, the interactions among the aromatic compounds may have enhanced overall biodegradation rates by allowing microbial growth instead of co-inhibiting biodegradation. There is enough evidence to support the conclusion that the pilot-scale reactor responded similarly to the lab-scale experiments previously reported for this design.     

Properties of phenol-removal aerobic granules during normal operation and shock loading

The physical structure and activity of aerobic granules, and the succession of bacterial community within aerobic granules under constant operational conditions and shock loading were investigated in one sequencing batch reactor over ten months. While the maturation phase of the granulation process began on day 30, the structure of microbial community changed markedly until after three months of reactor operation under constant conditions with a loading rate of 1.5 g phenol L⁻¹ day⁻¹. A shock loading of 6.0 g phenol L⁻¹ day⁻¹ from days 182–192 led to divergence of bacterial community, an inhibition of the biomass activity, and a decrease in phenol removal rate in the reactor. However, phenol was still completely removed under this disturbance. After the shock loading, the mean sizes of aerobic granules increased, and the activity of the microbial population within the granules decreased, although there appeared highly resilient for the dominant bacterial community of aerobic granules which mainly included β-Proteobacteria. Correlation analysis suggested that biomass concentration and biomass loading were significantly related to the community composition of aerobic granules during the whole operational period. The development of a relatively stable bacterial community in aerobic granules implied that those distinct dominant microbes in aerobic granules were favorably selected and proliferated under the operational conditions.     

Toluene removal from waste air using a flat composite membrane bioreactor

In this report, gaseous toluene biodegradation results in a flat composite membrane reactor inoculated with Pseudomonas putida TVA8 are presented. Preliminary abiotic experiments showed that transport of toluene through the membrane was linearly and negatively correlated with the gas residence time (tau). During a 339-day biofiltration experiment, the influence of gas residence time (2-24 sec) and mass loading rate (B(v); 10-483 g x m(-3) h(-1)) on the toluene elimination capacity was investigated. A maximum elimination capacity (EC(max)) of 397 g x m(-3) h(-1) was achieved at tau = 24 sec and B(v) = 473 g x m(-3) h(-1). Expressed per unit membrane area, the EC(m,max) was 0.793 g x m(-2) h(-1), which is five times higher than results obtained with other membrane bioreactor experiments in the same range of loading rates. At low gas residence times, reactor performance was limited by mass transfer. Toluene concentration profiles along the membrane were measured for several biotic and abiotic conditions. For inlet concentrations (C(in)) up to 1 g x m(-3), more than 90% was eliminated at 15 cm from the reactor inlet. For C(in) > 1.65 g x m(-3), longer membranes are necessary to obtain these high removal efficiencies.     

Performance of a horizontal-flow anaerobic immobilized biomass (HAIB) reactor and dynamics of the microbial community during degradation of pentachlorophenol (PCP)

The anaerobic biological treatment of pentachlorophenol (PCP) and methanol as the main carbon source was investigated in a horizontal-flow anaerobic immobilized biomass (HAIB) reactor at 30+/-1 degrees C, during a 220-day trial period. The reactor biomass was developed as an attached biofilm on polyurethane foam particles, with 24h of hydraulic retention time. The PCP concentrations, which ranged from 2.0 to 13.0 mg/L, were controlled by adding synthetic substrate. The HAIB reactor reduced 97% of COD and removed 99% of PCP. The microbial biofilm communities of the HAIB reactor amended with PCP, without previous acclimatization, were characterized by polymerase chain reaction (PCR) and amplified ribosomal DNA restriction analysis (ARDRA) with specific Archaea oligonucleotide primers. The ARDRA technique provided an adequate analysis of the community, revealing the profile of the selected population along the reactor. The biomass activities in the HAIB reactor at the end of the experiments indicated the development of PCP degraders and the maintenance of the population of methanogenic Archaea, ensuring the high efficiency of the system treating PCP with added methanol as the cosubstrate. The use of the simplified ARDRA method enabled us to monitor the microbial population with the addition of high concentrations of toxic compounds and highlighting a selection of microorganisms in the biofilm.     

Stability of the ANAMMOX process in a gas-lift reactor and a SBR

In the last years, the ANAerobic AMMonium OXidation (ANAMMOX) process has been put forward as a promising alternative to treat ammonium rich wastewaters. An ANAMMOX gas-lift reactor and a sequential batch reactor (SBR) were operated during around 200 days in this study, reaching nitrogen loading rates (NLRs) of 2.0 and 0.75 g l(-1) per day, respectively. The efficiency in the nitrite (limiting substrate) removal was 99%. The ammonium and nitrite influent concentrations were increased stepwise until biomass in the reactors started to float. These flotation events coincided with periods when the NLR exceeded the maximum specific ANAMMOX activity (MSAA) of the sludge. The MSAA, determined in batch experiments, was 0.9 and 0.44 g g(-1) per day for biomasses from the gas-lift reactor and the SBR, respectively. Flotation of the biomass occurred most likely due to a granule density decrease caused by dinitrogen gas accumulation inside the granules and an apparent breakage of the granules. Further research is needed to understand this phenomenon and to optimise the corresponding strategies to counteract the flotation.     

Hydrocarbon biodegradation in oxygen-limited sequential batch reactors by consortium from weathered, oil-contaminated soil

We studied the use of sequential batch reactors under oxygen limitation to improve and maintain consortium ability to biodegrade hydrocarbons. Air-agitated tubular reactors (2.5 L) were operated for 20 sequential 21-day cycles. Maya crude oil-paraffin mixture (13,000 mg/L) was used as the sole carbon source. The reactors were inoculated with a consortium from the rhizosphere of Cyperus laxus, a native plant that grows naturally in weathered, contaminated soil. Oxygen limitation was induced in the tubular reactor by maintaining low oxygen transfer coefficients (k(L)a < 20.6 h(-1)). The extent and biodegradation rates increased significantly up to the fourth cycle, maintaining values of about 66.33% and 460 mg x L(-1) x d(-1), respectively. Thereafter, sequential batch reactor operation exhibited a pattern with a constant general trend of biodegradation. The effect of oxygen limitation on consortium activity led to a low biomass yield and non-soluble metabolite (0.45 g SS/g hydrocarbons consumed). The average number of hydrocarbon-degrading microorganisms increased from 6.5 x 10(7) (cycles 1-3) to 2.2 x 10(8) (cycles 4-20). Five bacterial strains were identified: Achromobacter (Alcaligenes) xylosoxidans, Bacillus cereus, Bacillus subtilis, Brevibacterium luteum, and Pseudomonas pseudoalcaligenes. Asphaltene-free total petroleum hydrocarbons, extracted from a weathered, contaminated soil, were also biodegraded (97.1 mg x L(-1) x d(-1)) and mineralized (210.48 mg CO2 x L(-1) x d(-1)) by the enriched consortium without inhibition. Our results indicate that sequential batch reactors under oxygen limitation can be used to produce consortia with high and constant biodegradation ability for industrial applications of bioremediation.     

Phenol biodegradation by Pseudomonas putida DSM 548 in a batch reactor

Phenol biodegradation in a batch reactor using a pure culture of Pseudomonas putida DSM 548 was studied. The purpose of the experiments was to determine the kinetics of biodegradation by measuring biomass growth rates and phenol concentration as a function of time in a batch reactor. The Haldane equation μ=μ(m)S/((K(s)+S+S(2))/K(i)) adequately describes cell growth with kinetic constants μ(m)=0.436h(-1), K(s)=6.19mgl(-1), K(i)=54.1mgl(-1). These values are in the range of those published in literature for pure or mixed cultures degrading phenol.     

固定化微生物处理模拟污染地表水

以聚乙烯醇和海藻酸钠为包埋剂、驯化后的活性污泥为包埋菌剂,制备固定化微生物颗粒,其中包埋剂与包埋菌剂的比例为2:1。将该固定化微生物颗粒按20%的填充率装填到自制反应器中,用于处理模拟污染地表水,研究该固定化微生物的性能特点及其对模拟污染地表水的净化效果。结果表明:固定化微生物反应器的最佳水力停留时间为10h,最佳进水COD负荷为1.15～1.85g·L-1·d-1。在水温为20～29℃、溶解氧为3～4mg·L-1、水力停留时间为10h的条件下,当进水COD浓度为70.58～91.76mg·L-1、铵氮浓度为13.68～17.82mg·L-1时,COD去除率>62.3%,铵氮去除率>90.6%,表明固定化微生物能够有效地去除污染地表水中的COD和铵氮。     

Influence of substrate surface loading on the kinetic behaviour of aerobic granules

In the aerobic granular sludge reactor, the substrate loading is related to the size of the aerobic granules cultivated. This study investigated the influence of substrate surface loading on the growth and substrate-utilization kinetics of aerobic granules. Results showed that microbial surface growth rate and surface biodegradation rate are fairly related to the substrate surface loading by the Monod-type equation. In this study, both the theoretical maximum growth yield and the Pirt maintenance coefficient were determined. It was found that the estimated theoretical maximum growth yield of aerobic granules was as low as 0.2 g biomass g^–1 chemical oxygen demand (COD) and 10–40% of input substrate-COD was consumed through the maintenance metabolism, while experimental results further showed that the unit oxygen uptake by aerobic granules was 0.68 g oxygen g^–1 COD, which was much higher than that reported in activated sludge processes. Based on the growth yield and unit oxygen uptake determined, an oxidative assimilation equation of acetate-fed aerobic granules was derived; and this was confirmed by respirometric tests. In aerobic granular culture, about 74% of the input substrate-carbon was converted to carbon dioxide. The growth yield of aerobic granules was three times lower than that of activated sludge. It is likely that high carbon dioxide production is the main cause of the low growth yield of aerobic granules, indicating a possible energy uncoupling in aerobic granular culture.     

Effects of Phenol Feeding Pattern on Microbial Community Structure and Cometabolism of Trichloroethylene

Cometabolism of trichloroethylene (TCE) by phenol-fed enrichments was evaluated in four reactors with distinct phenol feeding patterns. The reactors were inoculated from the same source, operated at the same average dilution rate, and received the same mass of phenol over time. Only the timing of phenol addition differed. Reactor C received phenol continuously; reactor SC5 received phenol semicontinuously--alternating between 5 h of feed and 3 h without feed; reactor SC2 alternated between 2 h of feed and 6 h without feed; and reactor P received a single pulse every 24 h. The structure of the enrichments and their capacity for TCE transformation were analyzed. In long-term operation, reactors C and SC5 were dominated by fungi, had higher levels of predators, were more susceptible to biomass fluctuations, and exhibited reduced capacity for TCE transformation. Reactors P and SC2 were characterized by lower levels of fungi, higher bacterial biomass, higher concentrations of TCE-degrading organisms, and higher rates of TCE transformation. After 200 days of operation, rates of TCE transformation increased 10-fold in reactor P, resulting in TCE transformation rates that were 20 to 100 times higher than the rates of the other reactor communities. The cause of this shift is unknown. Isolates capable of the highest rates of TCE transformation were obtained from reactor P. We conclude that cometabolic activity depends upon microbial community structure and that the community structure can be manipulated by altering the growth substrate feeding pattern.