Effect of ferrous sulphate on nitrification during simultaneous phosphorus removal from domestic wastewater using a laboratory scale anoxic/oxic reactor

In the present study a laboratory scale anoxic/oxic reactor was used to remove the important eutrophication nutrients such as phosphorus and nitrogen from synthetic domestic wastewater. Phosphorus was removed through simultaneous precipitation and was carried out using the coagulant ferrous sulphate FeSO₄ · 7H₂O. Total phosphorus in the effluent was controlled to below 1 mg/l using a ferrous to phosphorus molar ratio of 2.1. pH after the addition of coagulant plays a major role in determining the molar ratio of the precipitant. Nitrogen was removed biologically in the anoxic/oxic system and the effect of simultaneous precipitation on nitrification and denitrification was investigated. The nitrification rate of the system remained unaffected during simultaneous precipitation and varied from 0.046 to 0.059 g N–NH₄ ⁺/g VSS/day. Denitrification was complete and was not affected by the coagulation process. The nitrogen removal efficiency varied from 78% to 85%. COD removal efficiency was not affected during simultaneous precipitation and was varied from 94% to 98%. The highly efficient nitrogen removal in the presence of simultaneous precipitant ferrous sulphate makes the process an ideal option for nutrient removal.     

Effect of alum on nitrification during simultaneous phosphorous removal in anoxic/oxic reactor

Phosphorus and nitrogen are the important eutrophication nutrients. They are removed in the anoxic/oxic reactor through simultaneous precipitation and biological nitrogen removal. The effect of alum a commonly used simultaneous precipitant on biological nitrification and denitrification are investigated in the present study. Simultaneous removal of phosphorus was carried out using the coagulant alum Al₂(SO₄)₃·14H₂O at 2.2 mol ratio. Before the start of simultaneous precipitation the nitrification rate of the A/O reactor was found to be 0.05 g N-NH₄ ⁺/g VSS/d. It starts to decrease with increase in coagulant dosage. The nitrification rate for alum dosage 97.13 mg/L was 0.38 g N- NH₄ ⁺/g VSS/d. There was no accumulation of nitrate in anoxic tank. The nitrogen removal efficiency of the reactor was affected and it fell from 88 to 78%. There was a slight decrease in effluent COD from 16∼20 mg/L to 8∼12 mg/L after the introduction of simultaneous precipitation into the reactor. The usage of alum as a simultaneous precipitant in the anoxic/oxic reactor was limited due to its inhibition on nitrification. Alum did not have any affect over denitrification process.     

Preliminary study of a suspended growth predenitrification system

A laboratory study has been conducted to obtained preliminary process information of a suspended growth Predenitrification (SGPDN)system. System performance was evaluated, in terms of chemical oxygen demand (COD) removal, NH(3)-N removal, system biomass yield and inventory, and effluent qualities, at different solids retention times (SRTs) and recycle ratios. Chemical oxygen demand removal in an SGPDN system occurs mainly in the anoxic reactor, which accounts for 94% of total COD removal. The overall COD removal rate is independent of recycle ratio (ranging from 2-5) used in this study; however, effluent COD increase with increasing recycle ratio. The observed anoxic and aerobic COD removal rates decrease with increasing SRT. The NH(3)-N removal in an SGPDN system is induced by two mechanisms: assimilatory NH(3)-N requirement for biomass production in the anoxic reactor and nitrification in the aerobic reactor. The observed anoxic NH(3)-N removal rate relates directly to the anoxic COD removal rate and agrees fairly well with the assimilatory NH(3)-N requirement theoretically predicted. The overall NH(3)-N removal rate is independent of SRTs and recycle ratios used in this study. Biomass yield in an SGPDN system occurs mainly in the anoxic reactor. However, uniform distribution of biomass throughout the entire system is obtained because of the high recycle rate used. The observed biomass yield (Y(O)) decreases with increasing STR. Tertiary treatment efficiency can be achieved in an SGPDN system. More than 90% reduction in feed COD., feed NH(3)-N, and NO(2) + NO(3)-N is obtained at all SRTs and recycle ratios used in this study. Higher MLVSS loading rates can be applied to a final clarifier without impairing its separation efficiency because of the excellent settleability of the Predenitrification activated sludge.     

Bacterioplankton Community Diversity in a Series of Thermally Stratified Lakes

Abstract The dominant members of the bacterioplankton community in a set of 10 small, thermally stratified lakes in northeastern Indiana were determined by denaturing gradient gel electrophoresis (DGGE) of a polymerase chain reaction amplified fragment of 16S rDNA. The variability in community composition was analyzed as function of vertical stratification (epilimnion vs metalimnion), time (July vs August samples), and geographical location. In 58 discrete samples, a range of 8–23 bands were detected (mean = 14, s.d. = 4). For all variables, sample pairs shared about 40–70% of bands. In comparisons between depth strata, pairs of oxic samples shared more bands than an oxic–anoxic pair. There was no obvious relationship between the geographical location of lakes (or their physical connection) and band sharing. Received: 4 March 1999; Accepted: 11 May 1999     

Effect of carbon source on biological nutrient removal in a sequencing batch reactor

Sequencing batch operation was used for nutrient (COD, NH4-N, NO3-N, PO4-P) removal from synthetic wastewater by using different carbon sources. Operation consisted of anaerobic, anoxic, oxic, anoxic and oxic (An/Ax/Ox/Ax/Ox) phases with durations of 2/1/4.5/1.5/1.5 h. Glucose, acetate and a mixture of glucose/acetate were used as carbon source to yield a COD/N/P ratio of 100/5/1.5 in the feed. Sludge age was kept constant at 10 days. COD, NH4-N, NO3-N and PO4-P removal efficiencies were maximum at the levels of 96%, 87%, 81% and 90% respectively, when a mixture (50/50) of glucose and acetate was used.     

Performance evaluation of an anoxic/oxic activated sludge system: effects of mean cell residence time and anoxic hydraulic retention time

A laboratory investigation has been undertaken to asses the effects of two operating parameters, mean cell residence time (MCRT) and anoxic hydraulic retention time (HRT), on the performance of an anoxic/oxic activated sludge system. The performance of the system was evaluated in terms of its COD, nitrogen, and biomass characteristics. An activated sludge system is capable of producing a better effluent, in terms of COD and nitrogen characteristics, when it is operated in an anoxic/oxic fashion. A longer MCRT and an adequate anoxic HRT are desirable in the operation of an anoxic/oxic activated sludge system. For the wastewater used in this investigation, the anoxic/oxic unit was capable of producing an effluent with the following characteristics when it was operated at MCRT = 20 days, total system HRT = 10 h, and anoxic HRT = 3-5 h: COD = 15 mg/L; VSS = 10 mg/L; TKN = 1.30 mg/L; NH(3) - N = 0.60 mg/L; and NO(2) + NO(3) - N = 5.0 mg/L. A uniform distribution of biomass is achievable in an anoxic/oxic activated sludge system because of the intensive recirculation/convection maintained. The provision of an anoxic zone in the aeration tank promotes a rapid adsorption of feed COD into the biomass without an immediate utilization for cell synthesis. This, in turn, results in a high microbial activity and a lower observed biomass yield in the system. A tertiary treatment efficiency is achievable in an anoxic/oxic activated sludge system with only secondary treatment operations and costs. A conventional activated sludge system can be easily upgraded by converting to the anoxic/oxic operation with minor process modifications.     

The influence of anaerobic pretreatment on the nitrogen removal from biosynthetic pharmaceutical wastewaters

The C:N ratio of the pharmaceutical wastewaters is usually suitable for a combination of the anaerobic pretreatment with the high COD removal and aerobic posttreatment with the efficient biological N removal. This kind of anaerobic-aerobic process was tested in semipilot scale by using a UASB reactor and an activated sludge system with a predenitrification (total volume 100 1). It was found that at a total HRT of 2.3 days an average of 97.5% of COD and 73.5% of total N was removed. The UASB reactor was operated at 30°C with a volumetric loading rate of 8.7 kg.m^-3.d^-1, the efficiency of COD removal was 92.2%. The processes, which take part in the biological removal of nitrogen, especially the nitrification, were running with lower rates than usually observed in aerobic treatment systems.Abbreviations AAO anaerobic anoxic oxic configuration - AOO anaerobic oxic oxic configuration - B _V volumetric organic loading rate (kg COD.m^-3. d^-1) - dB _x specific COD removal rate (mg COD. g^-1 VSS. d^-1) - DNR denitrification rate (mg N–NO₃. g^-1 VSS. h^-1) - E_COD efficiency of COD removal (%) - HRT hydraulic retention time (d) - NR nitrification rate (mg N–NO₃. g^-1 VSS. h^-1) - R recirculation ratio (%) - SBP specific biogas production (m³.kg^-1 removed COD) - SRT solids retention time; sludge age (d) - SS suspended solids (g.1^-1) - UASB upflow anaerobic sludge blanket reactor - VSS volatile suspended solids (g.1^-1)     

Simultaneous removal of carbon and nitrate in an airlift bioreactor

This paper presents the integrated removal of carbon (measured as chemical oxygen demand i.e. COD) and NO(x)-N by sequentially adapted sludge, studied in an airlift reactor (ALR). Simultaneous removal of COD and nitrate occurs by denitrification (anoxic) and oxidation (aerobic). Aerobic (riser) and anoxic (remaining part) conditions prevail in different parts of the reactor. Studies were carried out in a 42 L ALR operated at low aeration rate to maintain anoxic and aerobic conditions as required for denitrification and COD removal, respectively. The sludge was adapted sequentially to increasing levels of NO(x)-N and COD over a period of 45 days. Nitrate removal efficiency of the sludge increased due to adaptation and degraded 900 ppm NO(3)-N completely in 2h (initially the sludge could not degrade 100 ppm NO(3)-N). The performance of the adapted sludge was tested for the degradation of synthetic waste with COD/N loadings in the range of 4-10. The reduction of COD was significantly faster in the presence of NO(x)-N and was attributed to the availability of oxygen from NO(x)-N and distinct conditions in the reactor. This hypothesis was justified by the material balance of COD.     

Removal of carbonaceous and nitrogenous pollutants from a synthetic wastewater using a membrane-coupled bioreactor

Two modified Ludzack-Ettinger (MLE)-type membrane-coupled bioreactors (MBRs) were investigated in this study for the purpose of removing both nitrogenous and carbonaceous pollutants from a synthetic wastewater. During the first MBR experiment, removal efficiencies were high (>90%) for chemical oxygen demand (COD) and ammonia, but total nitrogenous pollutant removal efficiency was poor (~25%). Bacterial community analysis of ammonia oxidizing bacteria (AOB) by a nested PCR-DGGE approach detected two Nitrosomonas-like populations and one Nitrosospira-like population. During the initial portion of the second MBR experiment, COD and ammonia removal efficiencies were similar to the first MBR experiment until the COD of the influent wastewater was increased to provide additional electron donors to support denitrification. Total nitrogen removal efficiencies eventually exceeded 90%, with a hydraulic residence time (HRT) of 24 h and a recirculation ratio of 8. When the HRT of the MBR experiment was decreased to 12 h, however, ammonia removal efficiency was adversely affected. A subsequent increase in the HRT to 18 h helped improve removal efficiencies for both ammonia (>85%) and total nitrogenous compounds (~70%). Our research demonstrates that MBRs can be effectively designed to remove both carbonaceous and nitrogenous pollutants. The ability of the microbial community to switch between anoxic (denitrifying) and oxic (nitrifying) conditions, however, represents a critical process constraint for the application of MLE-type MBR systems, such that little benefit is gained compared to conventional designs.     

Influence of nitrate and COD on phosphorus, nitrogen and dinitrotoluene (DNT) removals under batch anaerobic and anoxic conditions

In this study, the effects of COD to NO(3)-N ratio in the feed on PO(4)-P removal was investigated. Maximum PO(4)-P uptake was obtained in the anoxic reactor when the COD to NO(3)-N ratios were between 2 and 3.75. With the influent COD of 800-1500 mg COD/L a total of the maximum removable PO(4)-P was 56 mg PO(4)-P/L through 20 days of anaerobic/anoxic incubation, indicating 98% P removal in the anoxic reactor. Similarly, for the COD to NO(3)-N ratios varying between 2 and 3.75 maximum denitrification was observed. Through anoxic operation the poly-P bacteria are capable of removing NO(3)-N using VFA, COD as carbon source and NO(3)-N as the electron acceptor after methanogenesis has been completed. High NO(3)-N concentrations stopped significantly the P uptake. A total of 97-99% dinitrotoluene removal efficiencies in the reactors containing COD to NO(3)-N ratio of 2 and 3.75 after 20 days of incubation period. For maximum NO(3)-N and PO(4)-P removals optimal COD to NO(3)-N ratios, COD and NO(3)-N concentrations were 2-3.75, 2000-4000 mg COD/L and, 800-1500 mg NO(3)-N/L, respectively.     

Simulated acid azo dye (Acid black 210) wastewater treatment by periodic discontinuous batch mode operation under anoxic–aerobic–anoxic microenvironment conditions

Treatment of simulated acid azo dye (C.I. Acid black 210) wastewater was studied in periodic discontinuous batch mode operation employing sequencing batch reactor (SBR) with suspended growth configuration under anoxic–aerobic–anoxic microenvironment. The performance of the reactor was evaluated at two organic loading rates (0.56 kg COD/m³-day and 0.75 kg COD/m³-day) with a total cycle period of 24 h [fill phase: 30 min; react phase: 23 h; settle phase: 15 min; decant phase: 15 min] at room temperature. The performance of SBR was assessed by monitoring both COD and colour (OD-617 nm) concentrations. Periodic discontinuous batch mode operation feasibility for dye colour removal along with simultaneous substrate (COD) removal. The performance efficiency of the system was found to depend on the operating organic loading rate. Induced anoxic microenvironment during cycle operation and persistent anoxic microenvironment in the internal layer of suspended biomass may be probable reasons for dye mineralization/reduction. Rapid startup period and non-inhibited performance at higher loading rate are some of the advantages observed in the SBR operation.     

Cloning and expression in Escherichia coli of a xylanase gene from Bacteroides ruminicola 23.

A gene coding for xylanase activity in the ruminal bacterial strain 23, the type strain of Bacteroides ruminicola, was cloned into Escherichia coli JM83 by using plasmid pUC18. AB. ruminicola 23 genomic library was prepared in E. coli by using BamHI-digested DNA, and transformants were screened for xylanase activity on the basis of clearing areas around colonies grown on Remazol brilliant blue R-xylan plates. Six clones were identified as being xylanase positive, and all six contained the same 5.7-kilobase genomic insert. The gene was reduced to a 2.7-kilobase DNA fragment. Xylanase activity produced by the E. coli clone was found to be greater than that produced by the original B. ruminicola strain. Southern hybridization analysis of genomic DNA from the related B. ruminicola strains, D31d and H15a, by using the strain 23 xylanase gene demonstrated one hybridizing band in each DNA.     

Cloning and expression in Escherichia coli of a xylanase gene from Bacteroides ruminicola 23

A gene coding for xylanase activity in the ruminal bacterial strain 23, the type strain of Bacteroides ruminicola, was cloned into Escherichia coli JM83 by using plasmid pUC18. AB. ruminicola 23 genomic library was prepared in E. coli by using BamHI-digested DNA, and transformants were screened for xylanase activity on the basis of clearing areas around colonies grown on Remazol brilliant blue R-xylan plates. Six clones were identified as being xylanase positive, and all six contained the same 5.7-kilobase genomic insert. The gene was reduced to a 2.7-kilobase DNA fragment. Xylanase activity produced by the E. coli clone was found to be greater than that produced by the original B. ruminicola strain. Southern hybridization analysis of genomic DNA from the related B. ruminicola strains, D31d and H15a, by using the strain 23 xylanase gene demonstrated one hybridizing band in each DNA.     

Succession of bacterial community structure and diversity in a paddy soil oxygen gradient

Cultivation-independent techniques were applied to assess the succession and phylogenetic composition of bacterial communities in a vertical oxygen gradient in flooded, unplanted paddy soil microcosms. Microsensor measurements showed that within 6 h of flooding, oxygen was depleted from 200 microM at the floodwater-soil interface to undetectable amounts at a depth of approximately 2 mm and below. The gradient was quite stable over time, although the oxygen depletion was less pronounced 84 days than 6 h after flooding. Community fingerprint patterns were obtained by terminal restriction fragment length polymorphism (T-RFLP) analysis from the oxic, transition, and anoxic zones of triplicate soil microcosms at 0, 1 and 6 h, and 1, 2, 7, 21, 30, 42, 84, and 168 days after flooding. Correspondence analyses revealed that T-RFLP patterns obtained using either community DNA or RNA were affected by time and oxygen zone, and that there was a significant interaction between the effects of time and oxygen zone. The temporal dynamics of bacterial populations were resolved more clearly using RNA than using DNA. At the RNA level, successional community dynamics were most pronounced from 1 h to 2 days and less pronounced from 2 to 21 days after flooding, for both oxic and anoxic zones. No effect of time or oxygen zone on the community dynamics was observed from 21 to 168 days after flooding. Dominant early successional populations were identified by cloning and comparative sequence analysis of environmental 16S rRNA and 16S rRNA genes as members of the Betaproteobacteria (oxic zone) and the clostridial cluster I (anoxic zone). Dominant late successional populations belonged to the Verrucomicrobia and Nitrospira (detected mainly in the oxic zone), and to the Myxococcales (detected mainly in the anoxic zone). In conclusion, the bacterial community developed through successional stages, leading at the RNA level to almost stable community patterns within 21 days after flooding. This principal finding, in combination with the phylogenetic identity of early- and late-appearing populations, suggests that the community dynamics can be explained by the principles of r- and K-selection.     

Alterations in the number of rRNA operons within the Bacillus subtilis genome

Deletions and additions of rRNA gene sets in Bacillus subtilis were observed by Southern hybridizations using cloned radiolabeled rDNA sequences. Of the ten rRNA gene sets found in B. subtilis 168M or NCTC3610, one was deleted in strains possessing the leuB1, ilvC1, argA2 and pheA1 mutations. Among EcoRI restriction fragments of genomic DNA products, a 2.9-kb 23S rRNA homolog was missing. In HindIII digest, both 5.5- and 5.1-kb hybrid bands were lost with 16S and 23S probes, respectively. Similarly, genomic DNAs digested with SmaI showed the absence of both 2.1- and 2.0-kb fragments that hybridized to 16S and 5S sequences, respectively, in wild-type genomes. In contrast, B. subtilis strain 166 and its derivatives displayed a gain of a 3.3-kb HindIII fragment homologous to 16S rRNA. Transforming the ilvC1 and leuB1 mutations into new genetic backgrounds revealed in some clones the concomitant introduction of the ribosomal defect. Transformations with the slightly heterologous donor DNA from strain W23 yielded some Leu+ and Arg+ transformants with altered hybridization patterns when probed with cloned sequences. We propose that the deletion of the rRNA operon occurred in the ilv-leu gene cluster of the B. subtilis genome as a result of unequal recombination between redundant sequences.     

A polymorphic DNA marker genetically linked to congenital malformations

Unusual restriction fragments were detected by DNA blot hybridization with PCNA (DNA polymerase-delta auxiliary protein) probe in one of seven cases of congenital malformations. Chromosomal in situ hybridization localized PCNA gene to region q31-35 of human chromosome 2. To discover the locus more closely associated with congenital malformations, a cloned DNA segment which has been mapped to chromosomal region 2q33-36 was tested for restriction fragment length polymorphisms (RFLPs) in these patients. The 2q33-36 probe hybridized with 2.1-kb, 1.9-kb and 1.7-kb fragments in ten normal control samples. In seven cases of congenital malformations examined, however, the band of 2.1 kb is absent in six cases and the band of 1.7 kb in one case. These results indicate that the locus closely linked to congenital malformations is present in the proximity of PCNA locus.     

Redox fluctuation structures microbial communities in a wet tropical soil

Frequent high-amplitude redox fluctuation may be a strong selective force on the phylogenetic and physiological composition of soil bacterial communities and may promote metabolic plasticity or redox tolerance mechanisms. To determine effects of fluctuating oxygen regimens, we incubated tropical soils under four treatments: aerobic, anaerobic, 12-h oxic/anoxic fluctuation, and 4-day oxic/anoxic fluctuation. Changes in soil bacterial community structure and diversity were monitored with terminal restriction fragment length polymorphism (T-RFLP) fingerprints. These profiles were correlated with gross N cycling rates, and a Web-based phylogenetic assignment tool was used to infer putative community composition from multiple fragment patterns. T-RFLP ordinations indicated that bacterial communities from 4-day oxic/anoxic incubations were most similar to field communities, whereas those incubated under consistently aerobic or anaerobic regimens developed distinctly different molecular profiles. Terminal fragments found in field soils persisted either in 4-day fluctuation/aerobic conditions or in anaerobic/12-h treatments but rarely in both. Only 3 of 179 total fragments were ubiquitous in all soils. Soil bacterial communities inferred from in silico phylogenetic assignment appeared to be dominated by Actinobacteria (especially Micrococcus and Streptomycetes), "Bacilli," "Clostridia," and Burkholderia and lost significant diversity under consistently or frequently anoxic incubations. Community patterns correlated well with redox-sensitive processes such as nitrification, dissimilatory nitrate reduction to ammonium (DNRA), and denitrification but did not predict patterns of more general functions such as N mineralization and consumption. The results suggest that this soil's indigenous bacteria are highly adapted to fluctuating redox regimens and generally possess physiological tolerance mechanisms which allow them to withstand unfavorable redox periods.     

para-Chlorophenol inhibition on COD, nitrogen and phosphate removal from synthetic wastewater in a sequencing batch reactor

COD, nitrogen, phosphate and para-chlorophenol (4-chlorophenol, 4-CP) removal from synthetic wastewater was investigated using a four-step sequencing batch reactor (SBR) at different sludge ages and initial para-chlorophenol (4-CP) concentrations. The nutrient removal process consisted of anaerobic, oxic, anoxic and oxic phases with hydraulic residence times (HRT) of 1/3/1/1 h and a settling phase of 0.75 h. A Box-Wilson statistical experiment design was used considering the sludge age (5-25 days) and 4-CP concentration (0-400 mg l(-1)) as independent variables. Variations of percent COD, NH4-N, PO4-P and 4-CP removals with sludge age and initial 4-CP concentration were investigated. Percent nutrient removals increased with increasing sludge age and decreasing 4-CP concentrations. Low nutrient removals were obtained at high initial 4-CP concentrations especially at low sludge ages. However, high sludge ages partially overcome the adverse effects of 4-CP and resulted in high nutrient removals. COD, NH4-N, PO4-P and 4-CP removals were 76%, 72%, 26% and 34% at a sludge age of 25 days and initial 4-CP concentration of 200 mg l(-1). Sludge volume index (SVI) also decreased with increasing sludge age and decreasing 4-CP concentrations. An SVI value of 104 ml g(-1) was obtained at a sludge age of 25 days and initial 4-CP of 200 mg l(-1).     

A new DNA marker, U15557, is linked to protease inhibitor and adenylate kinase-1 in the laboratory opossum, Monodelphis domestica

A 323-bp DNA fragment (U15557) was isolated, cloned, and sequenced after polymerase chain reaction (PCR) amplification from Monodelphis domestica genomic DNA. AHindIII restriction fragment length polymorphism was identified in this species using the U15557 PCR. fragment as a hybridization probe. DNA samples exhibited either a 6.4kb band, a 7.2 kb band, or both bands simultaneously. Behaviour of these two variants in family studies was consistent with codominant autosomal inheritance. Linkage between this marker and the loci encoding protease inhibitor (PI) and adenylate kinase 1 (AK1) was found in M. domestica.