Water Research 37 (2003) 2571–2582

Urea hydrolysis and precipitation dynamics in a urine-collecting system
Kai M. Udert, Tove A. Larsen*, Martin Biebow, Willi Gujer
Swiss Federal Institute for Environmental Science and Technology (EAWAG), Urban Water Management, P.O. Box 611, 8600 Dubendorf, Switzerland . Received 21 March 2002; accepted 21 January 2003

Abstract Blockages caused by inorganic precipitates are a major problem of urine-collecting systems. The trigger of precipitation is the hydrolysis of urea by bacterial urease. While the maximum amount of precipitates, i.e. the precipitation potential, can be estimated with equilibrium calculations, little is known about the dynamics of ureolysis and precipitation. To gain insight in these processes, we performed batch experiments with precipitated solids and stored urine from a urine-collecting system and later simulated the results with a computer model. We found that urease-active bacteria mainly grow in the pipes and are flushed into the collection tank. Both, bacteria and free urease, hydrolyse urea. Only few days are necessary for complete urea depletion in the collection tank. Two experiments with precipitated solids from the pipes showed that precipitation sets in soon after ureolysis has started. At the end of the experiments, 11% and 24% of urea was hydrolysed while the mass concentration of newly formed precipitates already corresponded to 87% and 97% of the precipitation potential, respectively. We could simulate ureolysis and precipitation with a computer model based on the surface dislocation approach. The simulations showed that struvite and octacalcium phosphate (OCP) are the precipitating minerals. While struvite precipitates already at low supersaturation, OCP precipitation starts not until a high level of supersaturation is reached. Since measurements and computer simulations show that hydroxyapatite (HAP) is the final calcium phosphate mineral in urine solutions, OCP is only a precursor phase which slowly transforms into HAP. r 2003 Elsevier Science Ltd. All rights reserved.
Keywords: Urine separation; NoMix; Ureolysis; Precipitation; Struvite; Calcium phosphate

1. Introduction The separate collection and treatment of urine is a possible way to reform today’s wastewater management. Since urine contributes most nutrients but also many problematic micropollutants to urban wastewater, urine separation has several benefits: it relieves conventional wastewater treatment, enables high nutrient recycling to

*Corresponding author. Tel.: +41-1-823-50-39; fax: +41-1823-53-89. E-mail addresses: udert@eawag.ch (K.M. Udert), larsen@eawag.ch (T.A. Larsen).

agriculture and facilitates the elimination of micropollutants [1]. Urine-separating toilets (NoMix toilets) and waterless urinals are used for collecting low diluted urine. First experiences show that blockages of the stench traps and pipes, caused by inorganic precipitates, are frequent and persistent [2]. They are a serious obstacle to a wide introduction of urine-collecting systems. In previous studies we have identified struvite, hydroxyapatite (HAP, Ca10(PO4)6(OH)2), and calcite (CaCO3) as precipitate compounds and we could show that microbial urea hydrolysis—also called ureolysis—triggers precipitation [3]. With the help of a computer model, we estimated precipitation potentials (PPs) as a function

0043-1354/03/$ - see front matter r 2003 Elsevier Science Ltd. All rights reserved. doi:10.1016/S0043-1354(03)00065-4

we focused on kinetics by examining ureolysis and precipitation in a urine-collecting system. both processes are negligible because the presence of efficient ureolysing bacteria is highly probable. availability of nitrogen sources. such as pre-existing crystals or foreign bodies and surfaces. we focused on the precipitation of calcium phosphate minerals and struvite. bacterial ureases appear to be located in the cytoplasma and are not deliberately excreted to the environment. pH increases during ureolysis. they also occur in urinecollecting systems. which is produced by several yeasts and algae [5].10]. especially when scales already exist.11]: dX 1=n p ¼ ksðIAP1=n À Ksp Þ . This process comprises the transport of lattice ions from the bulk to the crystal and their adsorption and dislocation on the crystal surface. However. much of the urease activity in soil is due to extracellular enzymes probably derived from plants and disintegrated bacteria [5. inhabit human intestinals and can cause urinary stone diseases [5–7]. kinetic constants can vary widely from species to species. Crystal growth describes the incorporation of solutes into the crystal lattice. Precipitation of phosphate minerals In this study. a positive SI is not sufficient for precipitation. In this study. mineral precipitation is only possible in supersaturated solutions. Ureolysis The enzyme urease (urea amidohydrolase) hydrolyses urea to ammonia and carbamate.8]. HAP precipitation is known to be preceded by the formation of other calcium phosphate minerals. We estimated the efficiency of urea-hydrolysing bacteria in the urine collection tank and in the pipes. Environmental conditions such as the If SI is positive. Several mechanisms have been proposed for describing surface-controlled crystal growth. A variety of eucaryotic and procaryotic organisms produce urease [5]. we elucidated the interaction of ureolysis and precipitation and we gained insight into the mechanism and sequence of precipitation. No evidence for substrate inhibition or allosteric behaviour has been detected [5. However. Since ureasepositive bacteria are ubiquitous. Denaturation of microbial urease occurs at pH values below 5. the urea concentration or the pH can regulate the urease synthesis. With the possible exception of urease from Helicobacter. In urine-collecting systems. and crystal growth [11. Therefore. Generally. The overall reaction can be written as follows: À NH2 ðCOÞNH2 þ 2H2 O-NH3 þ NHþ 4 þ HCO3 : ð1Þ Due to the ammonia release. There are several mechanisms for the regulation of urease synthesis. Most of them lead to an expression. The optimum pH of most microbial ureases is near neutrality. the occurrence of free ureases in urine-separating systems cannot be excluded. Therefore. even from strain to strain. They are widely distributed in the soil and in aquatic environments. urea decomposes non-enzymatically with a half-life time of 3. Michaelis–Menten kinetics can be applied to bacterial ureolysis. Nevertheless. the mineral can precipitate. because preliminary investigations have shown that precipitates from ureolysed undiluted urine consist of HAP and struvite. After nucleation the crystallites further increase to large crystals and particles by aggregation. while a negative SI indicates dissolution conditions. It provides information about the expected amount and composition of precipitates.M. Udert et al. dt ð3Þ . though bacteria are the most abundant.6 years at 38 C [9]. Since kinetic effects rather than solute–solid phase equilibria govern the initial crystallisation. In addition to urease. many bacteria synthesise urease constitutively [8]. a second enzyme is known to hydrolyse urea. in which crystal growth is a function of supersaturation [12. we also investigated the precipitation of commonly reported HAP precursors.8]. However. since an activation energy barrier prevents the lattice ions to aggregate as crystallites. the calculation of PPs considers only thermodynamic equilibria but no kinetic effects. The latter compound decomposes spontaneously to carbonic acid and a second molecule of ammonia [5]. crystallisation is usually controlled by surface processes. the composition of fresh precipitates can substantially differ from the model predictions.2572 K. The activation energy can be surmounted by high SIs and nucleation seeds. At supersaturation the ion activity product IAP of a mineral exceeds its solubility product Ksp [10]. The pH has low influence on the saturation constant KM of ureases but may strongly affect the specific activity. 3. coagulation. urea amidolyase. In toilet systems many nucleation seeds are present which promote the formation of crystallites. / Water Research 37 (2003) 2571–2582 of urine dilution and hydrolysed urea [4]. However. Furthermore. The PP is the mass concentration of precipitates when all solute and solid phases are in equilibrium. At low supersaturation. The saturation index SI provides a non-linear scale for supersaturation: SI ¼ log10 IAP : Ksp ð2Þ 2.

Cabrera. foreign ions can inhibit crystal growth by blocking growing sites. Direct transformation of ACP to HAP is also possible. thermodynamically more stable phases—such as HAP among calcium phosphates—are more sensitive to foreign substances [17]. with a short lifetime. We fixed the temperature to 2570. Heughebaert and Nancollas [13] found p ¼ 4 for the growth of octacalcium phosphate (OCP) at low supersaturation. / Water Research 37 (2003) 2571–2582 2573 where X is the mineral concentration in (mol lÀ1). when substituting phosphate [15].5 m were nearly horizontal including a 0. However. age 25–34). They concluded that OCP precipitation is governed by a polynuclear growth mechanism. In general. Struvite is less susceptible to inhibition. In complex solutions such as urine a variety of substances impede crystallisation. Dicalcium phosphate anhydride (DCPA. (Saltsjo-Boo. However. ACP is often the first phase. Gottingen. a magnetic valve was installed. CaHPO4 Á 2H2O) are precursors of HAP. we operated one reactor with fresh urine only.1 C. ACP formation can be skipped [13. Fresh urine was collected in bottles (six persons. Inhibition is well studied for calcite [14] but also for calcium phosphates. which were previously cleaned with 10% hypochlorite solution. 10 min) stored urine to the second reactor. Furthermore.. HAP is the most stable and the only one that will be found in urine solutions at solid–solute phase equilibrium [3]. and the sample interval was 1.1.g. The authors showed that under regular conditions and especially at elevated pH values struvite can precipitate but not newberyite. This value accords with the screw dislocation model by Burton. The experiments lasted 12 h. with ACP and OCP being the most likely in alkaline solutions. The inner diameter of the pipe and stench trap was 25 and 20 mm. 4. and urine sludge dissolved in filtered and centrifuged stored urine to the third reactor. and Frank [12]. [4] we give an overview of relevant complex formation reactions comprising ion pairs and inner-sphere complexes. Citrate. The three other reactors started with 250 ml fresh urine and 750 ml stored urine. and p a constant depending on the crystallisation mechanism.5 h. We added untreated stored urine to the first reactor.2. The resulting TSS concentration in the third reactor was 720 g TSS mÀ3. Concentrations in the solutions are given in Table 1. No detergents were used for toilet cleansing. Ca3(PO4)2). Neither citrate nor pyrophosphate inhibit struvite crystallisation strongly. At point 2. The high ionic strength as well as the organic and inorganic complexing agents reduce the concentration of free lattice ions substantially. and various macromolecules are important HAP inhibitors [15]. n the sum of all stoichiometric coefficients in IAP. In alkaline and slightly alkaline solutions.45 mm . [3].K. Each reactor was magnetically stirred. At low supersaturation. octacalcium phosphate (OCP. Sweden). magnesium. s a factor proportional to the total number of available growth sites.75 years. Both are magnesium minerals which have been detected in urinary stones. the value of p is typically 2 for many sparingly soluble salts. The total length of the pipe from toilet to collection tank was 3. and covered for preventing ammonia volatilisation. cellulose nitrate filters (Sartorius.15]. CaHPO4) and tricalcium phosphate (TCP. which had been running at EAWAG for 3. if crystalline nucleation seeds are abundant. which diverted flushing water.6 m long tube. especially HAP. Urea hydrolysis in the collection tank (tank experiment) To investigate urea hydrolysis in the collection tank we added fresh urine to stored urine from the collection tank.M. pyrophosphate. Measurements of precipitates and dilution factors are given in Udert et al. Udert et al. We ran four parallel glass reactors (volume 1 l).45 mm) and centrifuged (7000 rpm. We used medium hard drinking water for flushing [4]. Abbona et al. The toilet was a Type DS from Wost Man Ecology AB . E. They differed in the treatment of the stored urine. Materials and methods 4. Moreover. As reference. Ca8H2(PO4)6 Á 5H2O) and dicalcium phosphate dihydrate (DCPD. urine was diluted about four times and had a retention time of at least 3 weeks. HAP seldom precipitates directly from supersaturated solutions. In Udert et al. and were pressure-filtered through 0.6 m from the toilet. [18] investigated the conditions under which newberyite (MgHPO4 Á 3H2O) and struvite (MgNH4PO4 Á 6H2O) may occur in urine-like liquors. 4. Values behind the 7sign generally indicate population standard deviations. It is well established that amorphous calcium phosphate (ACP. respectively. However. but phosphocitrate is an efficient inhibitor [16]. The pipe material was PVC. which quickly converts to HAP. k the precipitation rate constant. followed by OCP. Germany). . Natural and technical aqueous systems are often supersaturated with respect to several calcium phosphate phases. of which about 1. which served as stench trap. carbonate can significantly increase the solubility of HAP. Ca3(PO4)2) have also been proposed as possible precursor but there is little evidence for its formation at ambient temperature.6 m. foreign ions can destabilise the crystal lattice when they are incorporated. In the collection tank. filtered (0. Investigated subject The subject of our investigation was a NoMix system.

700 ml solution. We name the corresponding reactors reactor horizontal and reactor vertical. sample surface in the pipe: 710 mm2. we performed a second experiment in which we added a 20 g N lÀ1 urea solution to untreated and to filtered stored urine.9 0.3.2574 K. we washed the collecting vessel with 10% hypochlorite solution before each sampling and centrifuged the urine (7000 rpm. The solid samples can be characterised as follows: Horizontal pipe: layer thickness: ca. All samples were stored at 4 C before analysis. chloride. Maidstone. and magnesium (72%) were determined with ICP-OES (Inductively Coupled Plasma-Optical Emission Spectrometer. TSS in the reactor: 810 g TSS mÀ3.M. / Water Research 37 (2003) 2571–2582 Table 1 Measured urine concentrations and literature values [4] Ureolysis in collection tank Fresh urine Mean Ammonia (g N m ) Urea (g N mÀ3) Phosphatea (g P mÀ3) Calcium (g mÀ3) Magnesium (g mÀ3) Sodium (g mÀ3) Potassium (g mÀ3) Sulphate (g SO4 mÀ3) Chloride (g mÀ3) Carbonatea (g C mÀ3) Total COD (g O2 mÀ3) pH (dimensionless) a b a À3 Ureolysis in pipes Fresh urine Mean CV% 2. Urea hydrolysis in NoMix pipes (pipe experiment) To determine the ureolysis activity in solids from the NoMix pipe we ran three reactors: one reference reactor and two reactors with different solid samples. calcium. [HCO3] and [CO3 ]. 1 mm.1 C. additionally 25 ml samples for TSS in the beginning and at the end of the experiments.2 386 8750 559 168 121 3730 2250 1350 5230 o5 9700 6. calcium (72%). sulphate.9 1. 2 mm all over the cross section. Chloride (precision of analysis: CV%=72%). TSS in the reactor: 804 g TSS mÀ3. magnesium. collected in bottles (six persons. for carbonate [H2CO* 3]. We took solid samples from a horizontal part (ca.7 3. channel height: ca. sample surface in the pipe: 1300 mm2. Again. and COD we conserved the samples with 65% HNO3.2 mm microfiltration unit (polypropylene membrane. . For analysis of phosphate. The reactors were 2 l Erlenmeyer flasks. We used stored urine from the same collection tank.5 m from the toilet). 4. Udert et al. The solution in each reactor was fresh urine.4. and phosphate (75%) were analysed with ion chromatography (column: IonPacs AS12A.3 Literature Urine Mean 480 7700 740 190 100 2800 2200 1500 3800 — — 6.4 2.6 1. covered for preventing ammonia volatilisation. Dionex Corporation.3 0.6 — 1720 73 76 28 1 837 770 292 1400 966 1650b 9. filled with ca. we filtered the urine with a 0. sample length: 20 mm. respectively. Vertical pipe: layer thickness: ca. CA.7 mm glass microfibre filters (Whatman. Potassium (73%). Sunnyvale. sulphate (72%). sample surface relative to the reactor volume (750 ml): 0.0b 254 5810 367 129 77 2670 2170 748 3830 — 8150 7. for being sure that all bacteria were separated. Microdyn. 1.1 4. so that the total ammonia concentration in the collection tank had risen from 1700 to 2700 g N mÀ3. 2. Total COD and pH in the collection tank origin from older measurements. and previously cleaned with 10% hypochlorite solution. total ammonia. sodium (73%). 30 mm.2 2. resulting in a sample pH below 2.7 1. 10 min). USA). UK). between the two tank experiments the urine dilution in the urine-collecting system had decreased. sample length: ca. To validate the first results.2 CV% 29 20 14 22 21 — — 29 — — — 8 Data range — — — — — 1800–5800 1300–3100 — 2300–7700 — — — Stored urine CV% 4.7 mÀ1. e. 8 mm.0 2À Including all system species. 4.g. Wuppertal. urea.5 h and the sample interval was 45 min. age 25–34).2 m from the toilet) and a vertical part of the pipe (ca. However. sample thickness ca. This time. This time.9 mÀ1. specific sample surface relative to the reactor volume (760 ml): 1. They were magnetically stirred.5 — 0. The experiments lasted 4.7 — — — — — 0.6 — 4.5 3. Sampling and analytical methods We took 25 ml samples for dissolved compounds. 2 mm. Germany) instead of centrifuging. The samples were pressure-filtered through 0. we fixed the temperature at 2570.8 1.

Greifensee. Furthermore. We performed simulations with different calcium phosphate phases and different p values. where urine is diluted up to 30 times. magnesium. except for potassium. The specific sample surface and the mass concentration of added solids were chosen according to the measured values. Loveland. It should be noted that Xstruvite is the actual mass concentration of newly formed struvite and not its activity. TCP. pH and concomitantly the proton concentration were fixed on the measurements. kstruvite the rate constant for precipitation (mol2 lÀ2 dÀ1). DCPD. rstruvite ¼ dxstruvite =dt ¼ kstruvite Xnucleate ðIAPstruvite À Ksp struvite Þp 1 þ signðIAPstruvite À Ksp  2 struvite Þ . Since the bacteria in our experiment had previously grown in pipes. potassium. We used the sum of squares of the weighted deviations w2 to assess quantitatively the consistence of 5. sodium. All analyses had mean recoveries of 100% 7confidence interval. This model is implemented in the computer simulation program Aquasim 2. As an example. ð4Þ where rurea is the rate of ureolysis (mol lÀ1 dÀ1). urea was hydrolysed to ammonia with urease (Merck 16493) according to EAWAG standard procedures. The precipitation rate constants were fitted on solute concentrations of calcium and magnesium in reactor horizontal and validated with measurements of reactor vertical. We determined total ammonia (72%) and urea (73%) photometrically (reaction with bromocresol purple) with flow injection analysis FIA (Ismatec AG. chloride. sulphate. Kleve. the rate of struvite precipitation was modelled as follows: . we applied least-square methods implemented in Aquasim. Weilheim. CellOx 225 WTW. we assumed constant bacteria concentration per unit surface. We used Eq. Before FIA analysis. The unit used for dissolved and solid components was mol lÀ1. Switzerland. and OCP. The accuracy of analytical methods was determined with recovery tests. COD (75%) was measured with HACH test tubes (HR. Udert et al. Preliminary studies showed that struvite and HAP are the sole minerals to be found in undiluted urine [3. TSS (75%) was measured as described in APHA standard methods [19]. citrate. In the simulation of ureolysis and precipitation in the NoMix pipes we used literature values for the complexing agents oxalate and citrate: 2. Glattbrugg.3  10À4 and 2. and pH. The p values were either 2 or 4. [4]. where the mean recoveries were 95%. carbonate. Germany). pH and oxygen were tracked with electrodes (combination pH electrode Mettler HA405DXK-S8/225. ð5Þ where rstruvite is the rate of struvite precipitation (mol lÀ1 dÀ1). CO. 0. Model To simulate the measurements of the pipe experiment we used a modified version of the precipitation potential model presented in Udert et al. Reference temperature is 25 C and activity coefficients are calculated according to the Davies approximation as cited in Stumm and Morgan [21]. Furthermore. Xnucleate the concentration of pipe sample (g TSS mÀ3) and sign(x) the sign of x (À1. Otherwise. To fit parameters. Precision of length measurement of the pipe samples was about 10%. kureolysis the zero-order rate constant of ureolysis (mol lÀ1 m dÀ1) and Ainoculate the specific surface area of pipe sample in the reactor (mÀ1). phosphate. We postulated that the crystallisation sites were proportional to the TSS concentration of the pipe sample and substituted the factor s from Eq. We introduced process kinetics for ureolysis and substituted the solute–solid equilibria with precipitation processes. (3) with XTSS : The last term in Eq. We fitted three parameters: the precipitation rate constants for struvite and calcium phosphate. Urea hydrolysis by urease usually follows Michaelis– Menten kinetics. Mettler-Toledo GmbH. þ1) (dimensionless). ammonium. HACH Company. oxalate. We did not simulate dissolution processes assuming that they were negligible in our experiments. USA). and the ureolysis rate constant. we postulated that struvite and one calcium phosphate mineral—HAP or a precursor— precipitates. (5) was introduced for preventing precipitation when the solution is not saturated. the activity of solids is set equal to unity [21]. sodium and COD. we assumed substrate saturation in undiluted urine and applied a zero-order kinetic: rurea ¼ Àkureolysis Ainoculate . when 1=3 1=3 the term (IAPstruvite 2Ksp struvite ) was negative and p was an even number.6  10À3 mol lÀ1. test tube 435. The model considers complexation and ionic strength effects. calcium. Input parameters are the total concentrations of urea. DCPA. / Water Research 37 (2003) 2571–2582 2575 1=3 1=3 Spectro Analytical Instruments.4].0 [20]. representing spiral growth mechanism or polynuclear growth mechanism. respectively [4]. Switzerland). precipitation would occur. respectively.M.K. Germany). We tested the following minerals as possible precursors of HAP: ACP. Therefore. Xstruvite the concentration of new struvite precipitates (mol lÀ1). Conventionally. (3) as basic approach for precipitation kinetics postulating that precipitation is controlled by surface dislocation processes.

6. / Water Research 37 (2003) 2571–2582 measurement and simulation [20]  n  X Smeas. Ammonia production in centrifuged stored urine indicated that free urease was present.i the ith measurement of fitted concentration (mol lÀ1). Smeas. when we related the rates to the wetted sample surfaces in the pipe.1 m vertical sections).i (mol lÀ1) and Si ðPfit Þ the calculated value corresponding to Smeas. A 14% of ureolysis activity was due to free urease.1 m3 stored urine). Lines are fitted with linear regression. the total ammonia concentration in the collection tank had increased from 1700 to 2700 g N mÀ3 due to lower urine dilution. The specific activities came to 49007600 g N mÀ2 dÀ1 in reactor vertical and 58007700 g N mÀ2 dÀ1 in reactor horizontal.2. we had to correct the measured total ammonia concentrations in the other reactors for determining the net total ammonia production by stored urine and urine sludge.2576 K. centrifuged urine. 3). We fitted the total ammonia concentrations with linear regression and obtained a ureolysis rate of 1820 g N mÀ3 dÀ1 (standard deviation7110 g N mÀ3 dÀ1) for untreated stored urine. The rates were 27707230 g N mÀ3 dÀ1 in untreated urine and 6307200 g N mÀ3 dÀ1 in filtered urine. we treated urine with filtration (0. smeas. In the pipe experiment. One indication is the elevated pH value (Table 1). Ammonia production in the first tank experiment.5 m horizontal sections and 2. 1). These values enabled us to estimate the urease activities in the investigated NoMix system. Fig. urease-active bacteria and free urease. ammonia was produced (Fig.18 g COD g TSSÀ1. The sum of total ammonia and urea remained constant in a range of maximally 72. We also detected ammonia formation in the reference reactor. The COD content of the urine sludge was 0. We investigated urease activity in the pipe of the NoMix system. the ammonia production was accompanied by a sharp pH increase (Fig. To determine the main cause of ureolysis we ran reactors with untreated stored urine. The pH kink at 7. In this experiment. 1.i i ¼1 ð6Þ where Pfit is the fitting parameter. which should not contain any bacteria. smeas. The average specific rate per surface is 54007900 g N mÀ2 dÀ1. Urine sludge appeared to contain no ureolysing bacteria.1. since rates in centrifuged stored urine with and without urine sludge were similar. Consequently. suspended bacteria were the main cause for urea hydrolysis. Ammonia production could sufficiently be fitted with linear regression. we diminished the concentrations by subtracting the measured values in the reference reactor in proportion to the volume of added fresh urine. the better the match of measured and simulated concentrations. respectively. 6. Consequently. .5% indicating that ammonia volatilisation was negligible. Results 6. In all three reactors. Hence. the fresh urine contained urea hydrolysing bacteria.i corresponded to the precision of analysis stated in the Materials and Methods section. Bacteria could either be suspended in the solution or settled to the bottom as part of urine sludge [3]. Here. The difference in the rates diminished. and 230740 g N mÀ3 dÀ1 for centrifuged urine with urine sludge. and centrifuged urine with urine sludge. In the meantime. About 960 g urea-N dÀ1 could be hydrolysed in the pipe (1. Ureolysis We expected two causes for ureolysis in the collection tank. Free urease was responsible for 23% of the ureolysis activity.M. and 180 g urea-N dÀ1 in the collection tank (filled with 0.i (mol lÀ1).i the standard deviation of Smeas. 2).2 coincided with the onset of struvite precipitation (Fig. The fit of the total ammonia concentrations resulted in ureolysis rates of 46107180 and 98607330 g N mÀ3 dÀ1 for the reactor vertical and the reactor horizontal.i À Si ðPfit Þ 2 w2 ðPfit Þ ¼ smeas. the composition of fresh urine corresponded well with literature values (Table 1). We performed a second tank experiment with stored urine. All rates were related to the volume of stored urine. 260730 g N mÀ3 dÀ1 for centrifuged urine. The two activities do not differ significantly. Udert et al.2 mm) for being sure that all bacteria were separated. Solution composition In the tank experiment. The smaller w2 . Only the phosphate concentrations differed significantly.

we had to determine which calcium phosphate mineral precipitated and which were the p factors of the rate equations. Measured (points) and fitted (solid lines) concentrations of dissolved components in reactor horizontal. 4. Dotted lines: 95% confidence intervals of model. Fig. To validate the fitting results. Dotted lines: 95% confidence intervals of model. . Precipitation of calcium was smaller. 6. respectively. we simulated the measurements in reactor vertical with the same three rate constants.K. 70% of dissolved calcium was incorporated into solids. These values agreed well with the measured values of 0.9 and 1. This value corresponded with the measured 810 and 804 g TSS mÀ3 for reactor horizontal and reactor vertical. We only changed the initial concentrations. Precipitation We observed strong precipitation in the pipe experiment (Figs. Here 90% of magnesium was eliminated in less than 1 h after struvite saturation had been exceeded. / Water Research 37 (2003) 2571–2582 2577 Fig. Measured (points) and simulated (solid lines) concentrations of dissolved components in reactor vertical. right side reactor vertical. In addition to the rate constants. Fig.3.8 mÀ1 for reactor horizontal and reactor vertical. 3 and 4). Validation of fit in reactor horizontal.9 and 1. and the sample surface. Left side reactor horizontal. more than 99% of dissolved magnesium precipitated during the whole experiment. Ammonia production in the pipe experiment. Udert et al. The ureolysis rate constant was proportional to the sample surface area. In contrast to ureolysis. Generally. In the reactor horizontal 1. 3. Dashed lines are fitted with linear regression. 2. precipitation rates were proportional to the mass concentration of added solids which we fixed at 800 g TSS mÀ3 for both reactors. Phosphate elimination was 44% in each reactor. In both reactors. We found good results with specific surface areas of 0. Rate constants for struvite and OCP precipitation were fitted to measured solute concentrations of calcium and magnesium.5 h were necessary for the same elimination.7 mÀ1 considering the measurement precision of 10%. We simulated precipitation and ureolysis in the pipe experiment by fitting their rate constants to measured solute concentrations of calcium and magnesium from reactor horizontal.M. respectively. precipitation was faster in the reactor vertical due to a faster pH increase by ureolysis. In both reactors.

Fitted for OCP precipitation with p ¼ 2 and 4: Fig.M. The best fit resulted with OCP as calcium phosphate mineral. 3 and 7). The latter was calculated with the precipitation potential model presented in Udert et al. calculation. which can be seen in Figs. In our model we usually considered two organic complexing agents. although we did not find references for such behaviour in literature. 6. 7). although OCP was supersaturated during the whole experiment. but wide for phosphate. the deviations were higher than for OCP (p ¼ 4) with w2 values of 593 and 442. the fitted model could still reproduce the course of measurements quite well (Fig. TCP. the fits were generally worse. and HAP precipitation. The fit with p ¼ 2 for OCP reproduced the sigmoidal run of the measurements badly and resulted in a w2 value of 410 (Fig. Struvite supersaturation therefore never reached high values (Fig. and p ¼ 2 for struvite (Figs. However. Fitted free calcium concentrations in reactor horizontal. However. Udert et al. The calculated . respectively. 5 and 6 and which is also indicated by the w2 values. 5). DCPD. In the initial simulations. The precipitation rate constant for struvite decreased 4 times and the one for OCP 4. respectively (graphics not shown). 5. The new fits were somehow worse with a w2 value of 548. we fixed the pH values on the measurements.2578 K. when pH was free for Fig. However. Calcium elimination proceeded slowly. For estimating their influence.2 Â 106 and 2. we neglected them in one simulation and refitted the precipitation rate constants for struvite and OCP (data not shown). citrate and oxalate. These minerals were partly undersaturated during precipitation. Once again. A refit of the precipitation rate constants resulted in a very good agreement for calcium and magnesium. / Water Research 37 (2003) 2571–2582 For fitting in validating. respectively. Fit of calcium concentrations in reactor horizontal assuming OCP. 9 illustrates that phosphate and magnesium elimination had already reached the values at equilibrium. The 95% confidence intervals of the model were narrow for magnesium and calcium. 3 and 4). We calculated the mass concentrations of OCP and struvite at the end of our experiment and compared them with the precipitation potential. We tested other calcium phosphate minerals and thereby used p values of 2 and 4. p ¼ 4 for OCP. the saturation maxima of DCPD and DCPA were not concomitant with the highest calcium precipitation rate (Figs. DCPA. All other processes and concentrations were unchanged. while the soluble calcium concentrations were still too high. Measurements and model predictions agreed well in both reactors with a w2 value of 367 in reactor horizontal. We also performed fits with p ¼ 4 for HAP or TCP. Saturation and concomitantly precipitation is highly influenced by complexing agents. Fits with HAP and TCP and p ¼ 2 predicted a steeper concentration decrease at the end of the experiment than observed (Fig. furthermore. (3) the precipitation rate is directly related to saturation. [4]. 6).8 Â 1017 m3 g TSSÀ1 molÀ3 l3 dÀ1 for struvite and OCP. Fig. but pH values still differed slightly (results not shown). The precipitation constants k came to 1. We assessed the final extent of precipitation by comparing the measured solute elimination with the theoretical elimination at solid–solute phase equilibrium. According to measurements and fits struvite precipitation set in abruptly after the saturation limit had been exceeded causing a sharp decrease in soluble magnesium and phosphate. The w2 values were 602 and 490. 8). so that saturation maxima and highest precipitation rates should coincide. according to Eq. and ACP were not suited for fitting the measurements. which means that complexation with citrate and oxalate reduced the precipitation rate by the same factors. we fixed the pH values on the measurements. which we attribute to the higher variation in the chemical analysis.8 times.

7.170. However.1. However. which corresponded to 87% and 97% of the precipitation potential. urea would be completely hydrolysed in the collection tank within little more than 1 day. respectively. the ureolysis rate in conventional collection tanks will be slightly lower. respectively. To classify the estimated urease activities. According to the measured urease activities. stored urine did not contain any significant amount of urea (Table 1). Nevertheless. Fig. the rates measured in our . Ureolysis In the investigated NoMix system. 8. Concomitantly. Accordingly. Urea was completely hydrolysed in the urine-collecting system. we can state that retention times of only few days are sufficient for complete ureolysis. a rough estimation shows that urea in restrained urine is completely hydrolysed: assuming that the channel of the investigated horizontal pipe is filled with undiluted urine. von Steiger et al. only about 11% and 24% of urea was hydrolysed. / Water Research 37 (2003) 2571–2582 2579 Fig. We cannot determine how much urea is hydrolysed in the pipes.M. right side reactor vertical.3 g N kg TSÀ1 dÀ1 (at 37 C) in a pasture soil near Zurich. Saturation indices during precipitation in reactor horizontal. Left side reactor horizontal. Calcium and magnesium concentrations and pH values from simulations without fixing pH to measurements. it takes only about 2 min for complete ureolysis. since data on the restrained urine volume are missing. [22] determined a urea hydrolysis of 3. 7. we can compare them with urease activities in agricultural soils. but urea was mainly hydrolysed in the collection tank. Discussion 7. total mass concentrations were 1370 and 1530 g TSS mÀ3 in reactor horizontal and reactor vertical. because the urine is not stirred. Udert et al. because most urine passed quickly through the pipe.K. Urease activity in the pipe was somewhat higher.

Hence. [23] reported that faecal bacteria added to stored urine died off more quickly at pH 8. A pH kink at about pH 7. / Water Research 37 (2003) 2571–2582 slightly inclined pipes most of the urea will be hydrolysed when the urine reaches the collection tank. although pH rose from 6. In both tank experiments. hence the urease activity per pipe surface was similar in both pipe sections. we found that ammonia production without bacteria is slower but still proceeds. and HAP are the crystalline minerals which occur in precipitates of ureolysed undiluted urine [3]. The calcium concentration was still considerably higher than at solid–solute equilibrium. This corresponds to measurements with Xray diffractometry. Since the phosphate concentration corresponded to its value at solid–solute equilibrium. OCP is only a precursor and transformation to HAP will follow. the increase of total ammonia could be well approximated with linear regression. In reactor horizontal and reactor vertical. On the one hand. The simulation of the precipitation potential give HAP and not OCP as calcium phosphate at solid–solute phase equilibrium. which indicated that struvite. 2). the specific urease activity differs between urine-collecting systems. struvite precipitation had already reached its final state. Ureolysis in pipes generally tends to increase with the retention time of the urine. respectively. urea hydrolysing bacteria probably die off in the collection tank. and are later flushed into the collection tank. further calcium phosphate formation was only possible by transformation of OCP into a calcium phosphate mineral. Therefore. The ureolysis rate correlated with the initial pipe surface. 87% and 97% of the precipitation potential were reached after 11% and 24% of urea had been hydrolysed.5 h after ureolysis started and at solid–solute phase equilibrium in reactor horizontal. This indicates that dissolved urease is present. We estimate that about 20% of the urease activity in the collection tank is due to free urease. respectively. Udert et al. Furthermore. Ureolysis and precipitation can be modelled based on first-order kinetics for urea hydrolysis and the surface dislocation approach for precipitation (Eq. On the basis of our results.9 and 10. Since we could not find any significant urease activity in the urine sludge.9 (reactor vertical). Consequently. 9. HAP formation may therefore set in as soon as struvite precipitation has reduced the concentration of soluble magnesium. experiments correspond to 310 and 60 kg pasture soil for the pipe and the collection tank (0.2. in very long and . where urease is released during cell lysis. Bouropoulos and Koutsoukos [24] could fit their experiments on spontaneous struvite precipitation with p ¼ 2: The good agreement with literature values supports our conclusion that OCP and struvite are the minerals that initially precipitate. However. Hoglund et al. As mentioned above.5 than at pH 6. 7. Heughebaert and Nancollas [13] already determined p ¼ 4 for OCP. Transformation of OCP to HAP has been described in literature.2 marks the onset of struvite precipitation (Fig. One reason could be the high pH in the . [25] showed that magnesium ions inhibit the crystallisation of HAP more strongly than the crystallisation of OCP. calcium phosphate formation was still in progress. ureolysis in completely stirred reactors with suspended bacteria is faster than in the pipes because of the increased contact of bacteria and urea. Fig. Hence. (3)). the kinetic constant that we fitted for the ureolysis rate is not generally applicable. In the pipe experiments.2580 K. Precipitation Our measurements confirm that microbial urea hydrolysis triggers precipitation. Although precipitation and transformation was still in progress at the end of the experiments.2 to 8.1 m3). the maximum mass concentration of precipitates was nearly reached. we assume that ureolysing bacteria grow in the pipes. we conclude that urease activity does not depend on pH in our system. Brown et al. Therefore. collection tank. [26] proposed the following reactions depending on the availability of calcium 1:25Ca8 H2 ðPO4 Þ6 Á 5H2 O-2Ca5 ðPO4 Þ3 OH þ 1:5H3 PO4 þ 4:25H2 O.0. Only little urea has to be hydrolysed for substantial precipitation. Fraction of precipitated solutes 4. Salimi et al.M. A direct correlation of pH value and increase of total ammonia was not visible. On the other hand. which has a higher Ca/PO4 fraction. The magnesium concentration corresponded well to its value at solid–solute equilibrium. The exponents in the precipitation rate equation p correspond with literature.

Udert et al. [9] Andrews RK. Estimating the precipitation potential in urine-collecting systems. strong struvite precipitation sets in.B. However. editors. In the collection tank. the pipes and in the collection tank causes fast precipitation of calcium and magnesium phosphates. Fixation of pH may be an appropriate means to consider their influence. 113–32. [7] Hasan HAH. Peters I. We assume that neglecting processes like carbon dioxide degassing or mineral dissolution is responsible for this deviation. Advances in inorganic biochemistry. Alder A. [5] Mobley HLT. as there are dilution of urine. In the reactors. Microbial ureases: significance. Marzilli LG. retention time of urine solution in the stench traps and pipes. In undiluted urine. because in our experiments calcium elimination did not reach its maximal value. Since hydroxyapatite (HAP) is the calcium phosphate phase at solid-solute phase equilibrium.M. Vol. 6. Maurer M. About 20% of the urease activity is due to free urease. Larsen TA.K. calcium. We proved that ureolysis triggers precipitation. and molecular characterization. editors. [10] Stumm W. p. While struvite precipitated completely until the end of the experiment. Island MD.59(3): 451–80. To find solutions for preventing blockages. However. In: Eichhorn GL. Commun Soil Sci Plan 2000. Biologically induced precipitation in urine-collecting systems.51(11):26–9. while OCP needs high supersaturation for substantial precipitation. In addition to inorganic precipitates foreign substances are an important cause for blockages of urine-collecting pipes. New York: Wiley. Eggen R. [4] Udert KM. magnesium. References [1] Larsen TA. Water Sci Technol. attachment of crystals on the walls. 1992. Wasser & Boden 1999. cohesion of minerals and erosion of scales. Hausinger RP. Presented at the Second IWA World Congress in Melbourne 2002. [6] Gleeson MJ. 8. / Water Research 37 (2003) 2571–2582 2581 Ca8 H2 ðPO4 Þ6 Á 5H2 O þ 2Ca2þ -2Ca5 ðPO4 Þ3 OH þ 3H2 O þ 4Hþ : ð7Þ The uptake of calcium seems to be the most likely transformation process. Philadelphia: W. Precipitation could be simulated with a dynamic computer model based on the surface dislocation approach. Swedish experiences with urine [2] Hellstrom separating systems. the suspended solids from the pipe surface not only increased ureolysis (see above) but also crystallisation. Infection stones. we investigated ureolysis and precipitation in a urine-collecting system. Griffith DP. Re-engineering the toilet for sustainable wastewater management. but high supersaturation is necessary for considerable OCP precipitation. lysis of urease-active bacteria sets urease free.31(15–16):2565–89. After precipitation had started. Blakeley RL. We found that urease-active bacteria primarily grow in the pipes. 245–83. 1990. it took only about 4 h for reaching the maximum mass concentration of precipitates in reactor vertical. For fitting and validating the measurements with our computer model. Long pipes may therefore enhance ureolysis. 2003. submitted for publication. Chemistry of the solid–water interface. maximum mass concentrations of precipitates can be reached in few hours. The long-term course of solute concentrations could eventually be simulated by including this transformation process in our model. the influence of these processes seems to be rather small. However. simulation could reproduce the course of pH. D. Amsterdam: Elsevier. regulation. OCP transforms later into HAP. [3] Udert KM. Environ Sci Technol 2001. all these influences have to be considered. Saunders Company. Muncke J. Microbiol Rev 1989. OCP formation was still in progress. Pak CYC. In: Resnick MI. Urolithiasis. Struvite precipitates immediately as soon as supersaturation is reached. Hausinger RP. Zerner B. Gujer W. urine retention times of only few days are necessary. Urea and urease. Johansson E. crystallisation in urine-collecting systems will be slower than in completely stirred lab-scale reactors. we had fixed the pH on measurements. while phosphate elimination already corresponded with the solid–solute phase equilibrium. Struvite precipitates earlier than OCP. . Gujer W. Molecular biology of microbial ureases. the maximum mass concentration of solids was nearly reached in both reactors indicating that only short urine retention times are necessary for maximum precipitate formation. The simulations resulted in struvite and octacalcium phosphate (OCP) as precipitating phases. Beside ureolysis and precipitation several other processes influence the occurrence and extent of inorganic blockages in urine-collecting systems. Ureolytic microorganisms and soil fertility: a review. Without pH fixation. Conclusions In this study.53(1):85–108. and phosphate (latter results not shown) quite well. p. For complete urea hydrolysis in the collection tank. A medical and surgical reference. 1984.35(9): 192A–7A. High urease activity in . Microbiol Rev 1995. Larsen TA. [8] Mobley HLT. Water Res 2003. but simulated and measured values differed more. which accords with literature. Soon after the solubility product has been exceeded.

2582 K. Spatial variation of urease activity measured in soil monitoring. Water Sci Technol 1998.88(12): 2478–81. 137–54. Formation and dissolution mechanisms of calcium phosphates in aqueous systems. . . Adv Colloid Interfac 1979. Boca Raton. Dubendorf. Calcified Tissue Int 1997. 3rd ed. Langmuir 1985. 73–82. Morgan JJ. [20] Reichert P.10:215–52. Smith M. Nancollas GH. Sallis JD. Stevens ED. [21] Stumm W. Hydroxyapatite and related materials. Stenstrom TA. . J Environ Qual 1996. Structure and chemistry of the apatites and other calcium orthophosphates. [17] Nancollas GH. Nancollas GH. Heughebaert JC. The growth of crystals in solution. p. Kinetics of crystallization of octacalcium phosphate. Koutsoukos PG.66(3):639–46. Nowack K. [13] Heughebaert JC. 1994. Switzerland: Swiss Federal Institute for Environmental Science and Technology (EAWAG). [14] Meyer HJ. Amsterdam: Elsevier. Washington DC: American Public Health Association and American Water Works Association and Water Environment Federation. [25] Salimi MH. p. New York: Wiley. / Water Research 37 (2003) 2571–2582 [19] APHA.38(6): 17–25. [15] Elliott JC. [12] Nancollas GH. Moore PB. 1996. J Cryst Growth 2000. editors. Tung TS. Crystal-chemistry of octacalcium phosphate.0—User Manual.61(3):216–22. In: Nriagu JO. Spontaneous precipitation of struvite from aqueous solutions. Constantz B.25(6):1285–90. Prog Cryst Growth Ch 1981. FL: CRC Press. Jonsson H. Evaluation of faecal contamination and microbial die-off in urine separating systems. 1995. Sundin A. Schulin R. [24] Bouropoulos NC. [18] Abbona F. 1998. [26] Brown WE. 1984.M. J Cryst Growth 1982. Berlin: Springer. [11] Nancollas GH.1(1):119–22. Boistelle R. [16] Wierzbicki A. . Zhang J. [23] Hoglund C.213(3–4):381–8. 19th ed. Udert et al. Aquatic chemistry. [22] von Steiger B. J Cryst Growth 1984. Aquasim 2. The nucleation and growth of phosphate minerals. Madsen HEL. Phosphate minerals.57(1):6–14. In: Brown PW. Crystallisation of two magnesium phosphates. Mathew M. Standard methods for the examination of water and wastewater. The influence of impurities on the growth rate of calcite. Sikes CS. Crystal growth of calcium phosphates in the presence of magnesium ions. struvite and newberyite: effect of pH and concentration. 4(1–2):59–87. Crystal growth and molecular modeling studies of inhibition of struvite by phosphocitrate. J Phys Chem 1984. . editors. 1994.

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