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Bacterial Competition in Activated Sludge: Theoretical Analysis of

Varying Solids Retention Times on Diversity
P.E. Saikaly and D.B. Oerther
Department of Civil and Environmental Engineering, University of Cincinnati, Box 210071, Cincinnati, OH 45221-0071, USA
Received: 19 February 2003 / Accepted: 19 June 2003 / Online publication: 3 May 2004

compete by lowering the shared pool of limited resources, as opposed to interference competition where
species compete by harming their rivals and by sequestering some of the resources for their exclusive use. Most
of these models were based on the classical ecological
principle of ‘‘competitive exclusion.’’ This principle,
which was pioneered by Lotka and Volterra [26, 48] and
first described mathematically by Voltera [49] and later
supported by several authors [1, 11, 28, 29, 36, 37], states
that the number of coexisting competing species, N, can
not exceed the number of growth-limiting resources
available to them (N £ K). This principle is the outcome
of competitive equilibrium or steady-state competition
(dX/dt = 0,where X is species abundance), where competition is allowed to proceed to equilibrium. It presumes
that increased competition for common resources causes
fewer numbers of species to coexist, leading to a predictable outcome where species that are strong competitors dominate. Many authors adopting this principle
showed theoretically and experimentally that the outcome of competition between two or more species
sharing a single limiting resource could be predicted
based on knowledge of Monod growth kinetics (half
saturation constant, Ks, of the growth-limiting resources
and maximum specific growth rate, rmax) and the species
death rate, b [5, 12, 23, 38]. Their results showed that
only one species is competitively dominant and the
remaining species are excluded. Similar results were obtained for several species competing for two resources
where eventually only two species can stably coexist [43,
Experimental and theoretical studies challenging the
competitive exclusion principle exist in the literature [2,
25, 39, 41, 45]. These studies rely upon nonequilibrium
conditions to promote species diversity by preventing
competitive equilibrium. An example of a process that
prevents competitive equilibrium from occurring is the
variability in resource supply ratios. Sommer [41] studied
the diversity of natural phytoplankton species in ‘‘che-


A mechanistic model for activated sludge sewage treatment was developed to predict exploitative competition
of six aerobic heterotrophic bacterial species competing
for three essential resources. The central hypothesis of the
model is that in a multispecies/limiting resource system
the number of coexisting bacterial species, N, exceeds the
number of limiting resources, K, available for them. The
explanation for this is that for certain species combinations, the dynamics of the competition process generate
oscillations in the abundances of species, and these
oscillations allow the coexistence of greater number of
species than the number of limiting resources (N > K).
This result is a direct contradiction of an existing activated sludge steady state competition theory, ‘‘the principle of competitive exclusion,’’ which states that the
competition process proceeds to equilibrium, allowing
only N £ K species to coexist. The model was used to
investigate the effect of varying solids retention times on
the diversity of species using the conventional, completely mixed activated sludge configuration. The results
of model simulations showed that for a certain range of
solids retention times (2.28–5.66 days) the competition
of six species for three essential resources produces
oscillations within the structure of the bacterial community allowing for the sustained growth of more than
three species on three resources.


Several mathematical models have been developed to
describe the effect of exploitative competition on the
coexistence of different species and thus, on species
diversity in natural ecosystems and ‘‘chemostat-type’’
bioreactors. Exploitative competition is when species
Correspondence to: D.B. Oerther; E-mail:


DOI: 10.1007/s00248-003-1027-6


Volume 48, 274–284 (2004)


Springer Science+Business Media, Inc. 2004

The rationale for modeling heterotrophic bacteria is that this ‘‘functional type’’ of organism is responsible for the removal of soluble. we hypothesize that the number of coexisting species. The ASM models describe microorganisms in activated sludge according to the average composition of three functional groups. when nutrients were provided in pulses. Activated sludge sewage treatment systems are engineered bioreactors used to remove organic substances and nutrients (nitrogen and phosphorus) from municipal wastewater. biomass is separated from the treated effluent through a biomass separator (e. OERTHER: IMPACT OF SRT ON 275 DIVERSITY mostat-type’’ bioreactors receiving continuous vs pulsed nutrients. . 14] describe processes occurring at the species level. would exceed the number of growth-limiting resources. A consortium of bacterial species is required to achieve the desired biological conversions. The results obtained with continuous nutrient addition showed that the community reached stable equilibrium (dX/dt = 0). The purpose of the clarifier is to selectively separate the retention of the biomass from the retention of the liquid.. a mechanistic model for activated sludge systems describing the competition of heterotrophic bacterial species for essential resources was developed. D. For the purpose of our simulations. and the number of species was equal to the number of limiting resources. Therefore. the number of coexisting species increased. the pulse nutrient condition corresponded to regular oscillations in some species. species of bacteria are ‘‘lumped’’ into one of these three functional groups. For clarity. Specifically.g. N. with multiple species and multiple limiting resources. The central hypothesis of our model is that in a complex system. autotrophs. Environmental engineers often use the term complementary nutrient in place of essential resource. a fraction of the biomass is intentionally removed from the system through the process of ‘‘wasting. Therefore. Methods The model developed in this study is based on mass balance equations for a conventional. 13. the Model Development. Huisman and Weissing [17–19] developed a series of models describing the competition of phytoplankton species for growth-limiting resources in aquatic ecosystems. and the loss of biomass was assumed to follow first-order decay and washout due to dilution. These oscillations prevented competitive equilibrium from occurring and allowed the coexistence of a greater number of species than the number of growth-limiting resources. From the clarifier. most of the models were developed to predict behavior in natural ecosystems or ‘‘chemostat-type’’ bioreactors. Although the studies described above show the importance of competition in predicting species diversity. In activated sludge systems. clarifier). and ASM3) [9. completely mixed activated sludge system (Fig. these models have not examined the impact of selective solids retention upon the competition process. or SRT. In the ASM models. Furthermore. None of today’s state-of-the-art activated-sludge models (e. However. K. The model was used to study the impact of the solids retention time (SRT) on the competition of heterotrophic bacteria for essential resources. we studied a broad range of SRT (0 to 30 days). namely heterotrophs. and the performance of these reactors largely depends on the bacterial community structure and their competition for different growth-limiting resources. understanding the dynamic behavior of different species competing for a variety of growth-limiting resources is expected to provide information useful for optimizing activated sludge systems. we have adopted the term essential resource throughout this manuscript.E. Essential resources obey Liebig’s law of the minimum where the concentration of the growth-limiting essential resource controls the overall growth of the organism. In the current study.g.P. Ranges of SRT for municipal activated sludge systems can vary from 1 day to 30 days with typical values for conventional systems of 2 to 5 days. Their work showed that competition could generate oscillations in the abundance of species. ASM2. For example. available to them (N > K). Their model was based upon a ‘‘chemostattype’’ system where growth was modeled using the Monod equation. ammonia and orthophosphate are examples of essential resources because they meet the requirement for nitrogen and Figure 1. Recently. Schematic of a completely mixed activated sludge sys- tem.. The process of wasting controls the solids retention time. which is the average time that biomass is retained in the reactor. Dashed lines designate the control volume used to generate the mass balance equations for essential resources and heterotrophic species. Essential resources fulfill metabolically independent requirements for growth. competition under certain operating conditions is expected to enhance species diversity. Therefore. phosphorus. Activated Sludge Model Number 1 [ASM1].’’ and the remaining biomass is recycled from the clarifier to the reactor. 1). SAIKALY.B. readily biodegradable organic pollutants typical of municipal sewage. and phosphorus-accumulating organisms (PAO).

(iii) the limiting resources are consumed by all of the different heterotrophic bacteria.90 0.20 2. and KKi is the half-saturation constant of species i on resource K. S2) change [3].10 0. Yji is the yield coefficient of species i for resource j.12 0. S1 is the growth-limiting resource for (S1.10 0.36 0. OERTHER: IMPACT OF SRT ON DIVERSITY Table 1. Xi is the concentration of species i.27 3. 24] and leads to a form of the overall growth equation where the specific growth rate is limited by one resource at a time (i.e.48 0.20 2.42 0.42 ð2Þ where K is the number of different essential resources.e. rmax1 = rmax2 = 1 d)1. ri will be equal to the lowest value in Eq. :::SK ÞXi V V ¼ Qin Sjo Qe Sj  Yji dt i¼1 for j ¼ 1.34 1. :::K ð3Þ where rmaxi is the maximum specific growth rate for species i. The model was developed to describe the competition of six groups of heterotrophic bacteria (e.36 0.SK) change).20 0.08 3.276 P.66 d)1). 34].88 0. ‘‘species’’) competing on three growth-limiting resources.30 1. (v) the hydraulic retention time (HRT) is kept constant at 0. To understand this equation. and the switching curve is a straight line passing through the origin with slope 0.96 3.. 19. and bi is the first-order decay rate for species i. (3):   rmaxi S1 rmaxi SK ..71 Resource 2 0.30 1.30 0. 31.12 0.18 0.96 3. growth is described by a switching function where the identity of the growth-limiting resource might change as environmental conditions (S1.17 0. consider an example of two essential resources.30 1. :::K ð1Þ   dXi V ¼ ri ðS1 . :::N Resource 1 2. Sj is the effluent concentration of resource j.26 2. and K2 = 0. Sjo is the influent concentration of resource j.17 0. :::SK ÞVXi  Qw Xi  bi Xi V dt for i ¼ 1.00 0. Qe is the effluent flow rate. Furthermore. 15.9. 1). Y Resource 3 0. In our model.33 1. This may or may not correspond to species as defined by 16S ribosomal DNA sequence comparisons [21.B. and (vi) the clarifier is ideal. We assumed that the specific growth rate on any essential resource is described by the Monod equation.17 0.88 2. The general forms of the differential mass balance equations for N heterotrophic species competing for K limiting resources are   N X dSj ri ðS1 .96 3.e.10 2.00 0.71 1.08 0.24 0.18 0. Next.88 0.14 0.14 0. a dilution rate of 1.07 selection of heterotrophic bacteria is consistent with the ecosystem of interest..9. Therefore.17 0. SAIKALY. assume that the maximum specific growth rate for both resources is the same. N is the number of different heterotrophic species.28 0. Model parameters Half saturation constant. i represents different 0.02 0.10 0. sources of carbon and energy) are present in excess.. (iv) competition is exploitative.21 0. whereas S2 is the growth-limiting re- . Although a thorough discussion of an appropriate definition of species is beyond the scope of the current work. Ks. 32] nor species as defined by operational taxonomic units (OTU) based on molecular fingerprinting assays [4. ri is the specific growth rate of species i.42 0. j represents different limiting resources. D.10 species. ::: ri ðS1 .08 Resource 3 0. mg/L Species 1 2 3 4 5 6 Resource 1 1.04 0. Qw is the waste flow rate (see Fig.6 days (i.55 0.00 0. we assumed that growth on essential resources obeys Liebig’s law of the minimum.46 0.14 0.20 1.g.. with assumed zero volume.17 0. In this case the equation of the switching curve becomes S2 = (K2/ K1)S1. In developing the model the following assumptions were made: (i) readily biodegradable substrates (i.E.77 3.43 Yield coefficient. The relationship between S1 and S2 where this equation is satisfied is called a ‘‘switching curve’’ because the identity of the growthlimiting resource switches when the environmental conditions (S1.76 0.47 1. (ii) oxygen is present in excess..14 0. namely activated sludge sewage treatment. Dual limitation by both resources is possible only under conditions where the growth rates are equal (r(S1) = r(S2)). :::SK Þ ¼ min K1i þ S1 KKi þ SK for i ¼ 1.24 0. K1 = 1. 17.33 0.21 0.14 0.46 Resource 2 0. The use of Liebig’s law of the minimum is well established [3.90 0.34 0.90 0. K1i is the half-saturation constant of species i on resource 1. species are defined as groups of like individuals that share a common set of kinetic and stoichiometric characteristics as defined in Table 1. S2) above and to the left of the switching curve. we feel that it is important to provide the ‘‘species concept’’ used throughout this manuscript. V is the volume of the completely mixed reactor.83 0. Qin is the influent flow rate.

predation of bacteria by protozoa. . 17.. The half-saturation constants and the yield coefficients were chosen to be unique for different growth-limiting resources (Table 1). and Ks change when the identity of limiting resource changes). Results Oscillatory Coexistence of Three Species on Three In this study we show Growth-Limiting Resources. The kinetic parameters (halfsaturation constant Ks. 2. heterotrophic bacteria in a conventional activated sludge system exhibit an oscillatory coexistence under a wide range of operating conditions. 2. The model was run for a period of 10.E. However. (A) Time course of competition for species 1. and 6. maximum specific growth rate rmaxi. 14]. Y. SAIKALY. Previous study [10] showed that the physiological state of an organism changes as the identity of the limiting resource changes (rmax. Oscillatory coexistence of three species on three essential resources at an SRT of 2. that for certain combinations of kinetic and stoichiometric parameters. and 6. model results showed no significant difference. 2 and summarized in the first column of Table 1. Model Parameters. and lysis) [8]. and 3. and the third is among species 1.000 days. 19]. and 5. Our model differs from the model described by Huismen and Weissing [17.66 d)1. All simulations were done using Berkeley Madonna Version 8.73 d and a dilution rate of 1.e. 13. we assumed a constant maximum specific growth rate that does not change when the identity of the limiting resource changes and a value of 1 d)1 was chosen for all species. The first competition is among species 1. The decay rate is a lumped parameter that includes several processes that lead to loss of biomass (i. D. The results of these simulations are shown in Fig. OERTHER: IMPACT OF SRT ON DIVERSITY 277 source for (S1. (C) Time course of competition for species 1. This assumption was based on previous work describing microbial growth on essential resources using Liebig’s law of the minimum and assuming a constant maximum specific growth rate [3. For simplification.0625 d. S2) below and to the right of the switching curve.1 [27] employing a fourth-order Runge– Kutta numerical approximation with a fixed time step of 0. (B) Time course of competition for species 1. we considered three examples of competition involving three species and three limiting resources. and 3. and yield coefficient Y) describing the heterotrophic species in this model were obtained from the literature [9. when we remove this constraint. To examine this phenomenon.25 d)1 was chosen for all species. endogenous respiration. or the mass of one cell. Instead the model sets Xi to be equal to zero when the mass of bacteria is <10)9 mg. 19].P. Figure 2. decay rate b. A first-order decay rate of 0. 2. and 5.0. 2. 3. and the patterns of oscillations were consistent throughout the simulation (results not shown).B. the second is among species 1. 3. in that it does not allow for continuity of Xi.

Species 1. SRT = 2.66 d)1.6 d). Oscillatory behavior was observed for a wide range of SRT values (species 1. and 3. OERTHER: IMPACT OF SRT ON DIVERSITY Figure 3. and 3 (Fig. SAIKALY. Species 1. 3. species 1. (i) It is already recognized in the literature that nonequilibrium conditions and oscillations allow the coexistence of more species than there are limiting resources [17–19. This is important because it is possible under these conditions to have coexistence of several species competing for the same resources. 2.3 mg/L) for resource 2 but can become limited by resource 1 or resource 3. respectively). (B) Oscillatory resource concentrations. 2. 41].0–4.3–3. and eventually excludes all other competing species that are limited by the same resource. and 3 establish a pattern of repeated oscillations due to competition. monopolizes the growth-limiting resource. SRT = 2.22–6 d.. and 6 were added to the system at t = 1000. we suggest that for oscillations to exist competition should be for at least three or more growthlimiting resources [17]. and 6 (Fig.3 mg/L) for resource 1 but becomes limited by resources 2 and 3. 2. 2. When the number of essential resources was reduced from three to two. (i) Oscillations are the result of nonequilibrium conditions that are generated by the competition process itself (i. 5. and 3 were added to the system at t = 0 days. and 3 for essential resources (Fig. SRT = 2. The cycle time for the oscillations in Fig. the oscillations that are caused by nonequilibrium conditions imposed by SRT create an opportunity to increase species diversity. 3A for species with kinetic parameters shown in Table 1 and SRT = 2. and 3 were large enough to create an opportunity for .3 mg/L) for resource 3 but becomes limited by resources 1 and 2.e. this pattern of transient dominance and oscillations did not occur.e. Previous competition theories. 3 and 5. Similar observations can be made for species combinations 1. 5. Two things can be deduced from these results. These limiting conditions lead to transient conditions where each species is dominant. species 4. based on equilibrium arguments. This result suggests that the amplitude of the oscillations generated by species 1. 2. Therefore. Species 4.73 d is 90 days. Species 3 is the best competitor (lowest Ks = 0. 2C). Oscillatory Coexistence of Six Species on Three The central hypothesis of Growth-Limiting Resources. 25. For example. and 5000 days. and 5 (Fig. consider the competition of species 1. It should be noted that adding all six species at t = 0 (results not shown) also resulted in the oscillatory coexistence of six species on three limiting resources. 47].73 d and dilution rate of 1. no species is capable of monopolizing the limiting re- source) for its limiting resource as each species switches the identity of its limiting resource through the course of competition. Three things can be deduced from the result. These oscillations occur because the species displace each other in a ‘‘round-robin’’ fashion [17.0 d.. 2B) and 1. 2000. our work is that oscillations in the abundance of species allow the coexistence of more species than growth-limiting resources. At later times. 2. (ii) No species is competitively dominant (i. 18. 2A) Species 1 is the best competitor (lowest Ks = 0. Therefore. (A) Oscillatory coexistence of six species on three essential resources at SRT = 2. To examine this phenomena.278 P. and 6 were able to coexist in the system because of the pattern of repeated oscillations created by the competition of species 1. consider the competition shown in Fig. 2. D. Species 2 is the best competitor (lowest Ks = 0. 3A). respectively.E. additional species are added (i. eventually lead to stable species composition where each species is competitively dominant on its growth-limiting resource. species 1.. This pattern of transient dominance repeats in an oscillatory fashion. 2A.e. 2 and 6.B. no external disturbance is necessary to generate oscillations).

The cycle time for the oscillations in Fig. (iii) Since these oscillations and coexistence are the result of nonsteady-state conditions. From an engineering standpoint. (B) Oscillatory coexistence of six species at SRT = 3 d. The competitive exclusion principle dominates for some values of SRT and is relaxed for other values of SRT.P. D.E. then one would predict that imposing non-steady-state conditions would result in oscillations of essential resources permitting a greater diversity of species to coexist on fewer limiting resources (Fig. 41] where variability in resource ratios creates non-steady-state conditions. 4B is 70 days compared to 90 days in Fig.000 days. Time course of competition of six species on three resources. and the only parameter that was varied was the SRT. our model maintained a constant concentration of resources in the influent. resulting in dynamic population composition and coexistence of the three functional group of organisms. 3B). and aerobic heterotrophs. at SRT value of 3 days. The coexistence of nitrifiers.0625 d. the effluent resource concentration is equal to the concentration inside the reactor. denitrifiers. (C) Steady– state coexistence of three species at SRT = 6 d. functional performance is based on steady-state conditions where the concentration of the essential resources are stable. and it produced the same pattern of cycles throughout (results not shown). SAIKALY. species diversity was evaluated using SRT values within the range of 0 and 30 days with an increment of 0. 4. (A) Steady-state coexistence of three species at SRT = 2 d.01 d. Figure 3B presents the effluent resource concentrations. In contrast. which means that increasing the SRT results in a concurrent decrease of cycle time. In the ASM models. Imposing non-steady-state conditions in activated sludge systems was shown theoretically and experimentally in a recent study using a completely stirred tank reactor (CSTR) with sludge recycle [16]. The model was run for a period of 20. This observation is important because it highlights a critical difference between our model and the ASM models. 3. To characterize this observation more thoroughly. (ii) The dynamics in the structure of the bacterial community impact the functional performance of the system as reflected by the oscillatory behavior in the concentrations of the essential resources (Fig. The results show steady state coexistence of three species on three growth-limiting resources (Fig.000 days simulation time with a fixed time step of 0. we have oscillatory coexistence of six species on three growth-limiting resources (Fig. A total of 3000 runs were conducted with 20. the impact of SRT on Figure 4. The results of model simulations at SRT = 2. was possible because the activated sludge reactor was exposed to sinusoidal oscillations of influent DO concentrations. 3B). 4A and C) at SRT values of 2 and 6 days. OERTHER: IMPACT OF SRT ON DIVERSITY 279 other species to coexist even at t = 0 days. This phenomenon is similar to the resourceratio theory [39. 4B). Spe- . and 6 days are shown in Fig. For completely mixed activated sludge systems. 3A.B. which promotes the coexistence of more species. the next step is to determine the operating conditions that trigger these oscillations and cause increased species diversity in activated sludge systems. whereas. Effect of SRT on Species Diversity. competing for the limiting resource dissolved oxygen (DO).

387 1. species 1. and in this case it is species 6 (Table 2).572 1. then the number of equilibrium solutions that satisfy Eq.647 will allow species 1 to grow. D.94 1.22 2. 4) [8]. For every SRT value examined in the range 1.72 d. SRTmin of different species for different limiting re- sources SRTmin (d) Species 1 2 3 4 5 6 S1 (mg/L) S2 (mg/L) S3 (mg/L) 1.472 1. Comparing the results of model simulation with Table 3.5 and 1.67–30 d the ‘‘competitive exclusion principle’’ dominates.647 and 1.5 2.436 1.21 2. we can see that SRT values in the range 0 to 1. Similar arguments apply to other SRT ranges where competitive exclusion dominates. Environmental engineers design activated sludge systems based on SRTmin (Eq.94 d.614 1.27 and 5.34–5.498 1. respectively. because Sio available for each species is lowered by the presence of other species competing for the same resource.392 1. 2000.463 1. and 6 are limited by S1. only one species can stably coexist. respectively (Table 2). This result is due to equilibrium conditions.280 P.521 1.28–2.33 4. and hence they both wash out. A possible explanation for such behavior is that the SRTmin required for the organisms to grow in a multi-species/resource system is greater than that predicted by Eq. For example.424 1. 5. Values of Ks were obtained from Table 1.660 1. if Eq. OERTHER: IMPACT OF SRT ON DIVERSITY Table 2. SRTmin ¼ Ks þ Sio Sio ðrmaxi  bi Þ  Ks bi ð4Þ where SRTmin is the minimum solids retention time and Sio is the influent resource concentration.B. 4.473 From these results.95–2. At SRT values between 0 and 1. and 6 were added at times 1000. Similarly. From the model simulations in the SRT range from 0 to 1. 2. then the number of species that can coexist cannot exceed the number of limiting resources available to them. and 5000 days.23 2.49 d.95–2. Table 2 is a summary of the simulation output showing coexistence (+) and washout ()) of six heterotrophic species competing for three growth-limiting resources at various SRT values.27 2. In other words.614 d. and only N £ K species coexist where K in our case is 3 (Table 2). At SRT values in the ranges 1. and the corresponding SRTmin are shown in bold (Table 3). and their corresponding SRTmin as determined by Eq. SAIKALY.72 1. any value of SRT that is greater than 1. any value of SRT that is greater than 1. Table 3.73–1. Therefore. (4).5–1. However. 2. and 3 were added at time zero.614 d will allow species 2 to grow. model results show that species 1 and 2 are limited by S1.517 1. Therefore. (4). we observed washout of all species because the maximum net specific growth rate is exceeded by the washout rate.73–1.67–30 d changes even though competitive .67–30 Species 1 Species 2 Species 3 Species 4 Species 5 Species 6 ) ) ) + + ) ) ) + + + + ) ) ) ) ) ) ) ) ) + + + ) + ) + + ) + ) + + + + ) ) ) ) ) ) ) ) ) + + ) ) ) ) ) ) ) ) ) + + ) ) ) ) + + ) + ) + + + + ) cies 1. their SRTmin for growth should be greater than that predicted by Eq. (1) and Eq. and hence they all wash out. (4) are 1.E. since we have K limiting resources.521 1. (2) at equilibrium with Xi > 0. whereas species 4. For operation at any SRT value that is lower than SRTmin.2 and 5.450 1. (2) are solved simultaneously for equilibrium (dSj/dt = 0 and dXi/dt = 0). whereas species 3 and 5 are limited by S2. since both species share the same resource (S1).24–2. (4) would not cause washout of the species from the system. Coexistence and washout of six species competing for three essential resources at various SRT SRT values (d) 0–1.647 1.49 1. Another deduction from Table 2 is that the identity of the competitively dominant species in SRT ranges 1. the model results show that we have only one steady-state value of resource concentration that satisfies Eq. However. traditional engineering theory would argue that an SRT value that is greater than SRTmin calculated by Eq. Table 3 presents the SRTmin of the six species when each exists alone and is limited by only one resource.73–2.49 d. at SRT values between 1.49 d are lower than the SRTmin for each species (bold numbers). species are removed from the system because of washout.372 1.2 2.51–4. For example.377 1.66 5. this is not the case in a multispecies system. (2) with Xi > 0 cannot exceed three.

Taken together.38 1. Discussion Our work had two overlapping goals. The recent work of Huisman and Weissing [17–19] is relevant to the present study. model simulations showed that N could be greater than K in some cases because of nonequilibrium conditions and oscillations in species concentration. we showed that under certain . respectively (Table 4).70 3. (7) in Eq.24 0.B. We ran a variety of simulations using different values of rmax (2 d)1). (2) at steady state (Eq.21 0.00 2. The second goal was to use this model to study the effect of operating parameters. The three steady-state values at SRT = 2 days correspond to species 1. values in the second column of each resource). (6): ri ¼ 1 þ bi SRT ð6Þ Substituting Eq. (2) at equilibrium with Xi > 0. Instead.00 2.90 3.02 2.64 0. :::N ð5Þ Substituting V/SRT for Qw and solving Eq. This observation may have important implications for ‘‘real-world’’ situations. As described by Sykes.90 3. then a nearly complete ecosystem model of the activated sludge process is computationally feasible. we get: O ¼ ri VXi  Qw Xi  bi Xi V for i ¼ 1. This implies that the impact of SRT on species diversity is a general phenomenon and not simply limited to the values of the kinetic parameters (Table 1) selected for these simulations. 5). SAIKALY. 2. The bold numbers in Table 4 correspond to the steady-state resource concentrations satisfying Eq.00 1. This is attributed to the fact that the steady-state resource concentration varies according to the SRT value (Eq.54 0. many environmental engineers approach activated sludge systems with the following belief [42]: ‘‘The number of coexisting species in an activated sludge process equals the number of limiting resources available to them and if all that is required is the solution of an N by N matrix (species by substrate).66 d showed that the competition of six heterotrophic species for three growth-limiting resources produced oscillations within the structure of the bacterial community allowing for the coexistence of more than three species on three resources (Table 2).06 3.74 0. D. The cycle time for the oscillations in Fig. and 6.’’ However.E.73 0. is not the only mechanism dictating the outcome of species competition for limited resources in activated sludge systems. the three steady-state values correspond to species 1.21 0.28–5. OERTHER: IMPACT OF SRT ON 281 DIVERSITY Table 4.64 0.28 2.24 0.31 0. whereas. the hypothesis proposed by Sykes is based on equilibrium arguments and the notion of competitive exclusion.70 0.70 0.55 0. (5) for r yields Eq.21 0. then reactors should be operated for a period greater than 30 days to experimentally observe oscillations.02 3.21 0. First. Steady-state resource concentration at SRT = 2 and 6 days SRT ¼ 2 days Species 1 2 3 4 5 6 SRT ¼ 6 days S1 (mg/L) S2 (mg/L) S3 (mg/L) S1 (mg/L) S2 (mg/L) S3 (mg/L) 3. 3. The cycle time has decreased to 30 days because of the higher values of rmax and Ks.33 0. (3) is solved for equilibrium.64 0.73 0. Two other interesting results are obtained from the model simulations. For example. Table 4 presents steady-state resource concentrations for the six species at SRT = 2 and 6 days using Eq.71 0.76 exclusion dominates in both ranges. Ks. The results of the model simulations suggest that the competitive exclusion principle. if the structure of the bacterial community in activated sludge systems is dynamic in nature and operates according to the conditions described in our model. on the competition process and species diversity. This equation is derived by solving Eq.71 0. 5B is 30 days.12 1.71 0. 5). for SRT = 6 days. (7). When Eq. and the results show similar trends (Fig. (1) and rearranging gives: S¼ Ks ð1=SRT þ bi Þ rmaxi  ð1=SRT þ bi Þ ð7Þ To better understand this point.06 2. They showed that nonequilibrium conditions and oscillations in the structure of microbial communities allow the coexistence of more species than there are limiting resources. in particular the effect of SRT. consider SRT values of 2 and 6 days. and Y (Table 1. This result is a direct contradiction of the competitive exclusion principle.90 3. 7). which states that N species must be equal to or less than K growth-limiting resources. The first goal was to derive a mechanistic model that could describe competition of heterotrophic species for growth-limiting resources in activated sludge systems. Simulations at SRT values between 2.and 3. the results of these studies are significant because they directly contract a long-held notion of microbial diversity in activated sludge systems.P.

(B) Oscillatory coexistence of six species at SRT = 3 d. and sameness of the identity of community biomass over temporal scales [46]. or hydraulic configuration of the aeration tank) can be used to enhance species diversity. If the diversity–stability hypothesis developed in these studies of macroecological systems applies to activated sludge systems. 46]. Second.e. This theoretical observation is supported by several recent studies. resilience (rate of recovery after disturbance). then we expect systems with higher diversity to better maintain performance when exposed to environmental perturbations (e.5 d. The controlling factor for the absolute level of species diversity was the P. Fernandez et al. OERTHER: IMPACT OF SRT ON DIVERSITY SRT. One example is the study done by Kaewplpat and Grady [20] where they examined the structure of the bacterial in a laboratoryscale sequencing batch reactor operated using denaturing gradient gel electrophoresis (DGGE) targeting 16S ribosomal RNA. The DGGE data showed that the bacterial community was highly dynamic. Thus. D.. These two observations have significant applications in design and modeling of activated sludge systems.282 Figure 5. [7] studied the community dynamics in a functionally stable methanogenic reactor over a period of . and this dynamic behavior was observed within the initial 17 days of reactor operation and continued throughout the experiment. we showed that this dynamic nature arises from the natural process of competition for essential resources. operating conditions competition for growth-limiting resources in activated sludge results in oscillations in the abundances of species.. toxic shock loads). Our result is novel for activated sludge models because traditional models do not include competition within functional groups. The first significant application is that the model could be used as a design tool to predict which engineering control strategies (i. The importance of species diversity was shown in a recent study examining toxic loads of mercury in bioreactors [6]. (A) Steady-state coexistence of three species at SRT = 1. Time course of competition of six species on three resources. Stability can refer to resistance to disturbance. This is important because both laboratory and field studies showed that diversity (species richness) is positively related to ecosystem stability [35. Furthermore. nor do they relate competition to the diversity of the microbial community in activated sludge. we showed that these oscillations allow the coexistence of more species and hence can increase species diversity. The results of the study showed that diverse biofilm communities demonstrated enhanced resistance to mercury toxicity as compared to monoculture biofilms. which show experimentally that the microbial community composition is dynamic. an increase in species diversity may increase the chance of obtaining species with different complementary physiological traits that are better adapted to handle specific environmental perturbations. HRT.B. Another significant application of the model is that for the first time we showed theoretically that the bacterial community structure in activated sludge system is dynamic. SRT.E. SAIKALY.g. Our results predict that there are situations where dynamic microbial communities are the norm and ‘‘steady-state’’ conditions do not exist. Studies using anaerobic reactors have also shown that the structure of bacterial communities is dynamic. (C) Steady-state coexistence of three species at SRT = 6 d.

a competition model that takes into consideration the difference in death rate between different bacteria (those with adaptive starvation response and those with no adaptive mechanism) could better describe the impact of decay rate on competition. Therefore. D. calcyflorus): An experimental test of a mechanistic model. New York 2. DB. The theoretical results of the current model and the results of the experimental studies discussed above suggest that microbial communities in aerobic and anaerobic bioreactors are dynamic. 13. Canstein. The current model assumes that the influent wastewater is composed of essential resources with bacterial species competing for these resources.’’ However. a more complete model should include multiple functional groups of organisms including aerobic and denitrifying heterotrophs. In a similar study. activated sludge systems are designed to remove nutrients as well as organics. RA. Blackall. Appl Environ Microbiol 68: 2829–2837 . Furthermore.P. Boraas. HV. 14]. Armstrong. and pH) over a 2-year period was dynamic. The reason we made this assumption is that most models of activated sludge wastewater treatment use a constant value for the decay rate of all heterotrophic organisms [9. In conclusion.E. namely the SRT. This reduced death rate allows these cells to be at a competitive advantage compared to cells with higher death rate. There are other design and operational parameters that can be modified. the mechanistic model that we developed to predict exploitative competition of six aerobic heterotrophic bacterial species competing for three growth-limiting resources showed that the competitive exclusion principle is not the only outcome of competition in activated sludge wastewater treatment systems. PL. in a recent study [22] it was shown that cells that undergo a starvation response during substrate pulse leading to periods of substrate limitation and hence low growth rate have a reduced death rate compared to cells that do not undergo a transition from active to dormant state. The results of their study showed that the bacterial community structure was dynamic and followed a chaotic behavior. Future model testing should be used to develop a ‘‘rank order’’ of the effectiveness of different operational parameters for affecting the diversity of the bacterial community in activated sludge systems. Zumstein et al. namely the ‘‘aerobic heterotrophs. Kelly. SAIKALY. the model was developed to study the competition of species within one functional group of microorganism. J Plankton Res 12: 77–87 6. [50] showed that the bacterial community structure in an anaerobic digester operated under constant environmental conditions (feeding. BC. JB (1990) Resource competition between two rotifer species (Brachionus rubens and B. may need to be modified to include non-steady-state conditions as well as competition and the dynamic behavior of the microbial community. Department of Civil and Environmental Engineering. Li. Baltzis. 2. including reactor configuration (plug flow versus complete mixed) and hydraulic retention time (HRT). and 3).. diversity is enhanced.28–5. Bond. Biotechnol Bioeng 31: 75–86 4. nitrifying autotrophs. LL.g. However.. P (1998) The use of 16S rDNA clone libraries to describe the microbial diversity of activated sludge communities. by the process of competition for essential resources as described in our model. Seale. Although the model provides a testable hypothesis for the impact of SRT on bacterial diversity in activated sludge systems. amino acids and ammonia are substitutable resources because they are both used as a source of nitrogen by aerobic heterotrophs). Therefore. Effects of competition in the presence of higher trophic levels.B. influent wastewater at a real wastewater treatment plant contains a variety of resources including essential and substitutable resources (e. We are also grateful for the helpful comments of two anonymous reviewers. D. We also showed that SRT is a key parameter in controlling competition and hence diversity in activated sludge systems. John Wiley & Sons. I (2002) Species diversity improves the efficiency of mercury-reducing biofilms under changing environmental conditions. Y. McGehee. temperature. Acknowledgments The authors gratefully acknowledge financial support from the National Science Foundation (BES 0116912) and the University of Cincinnati. we suggest that specific operating parameters for bioreactors affect microbial diversity by affecting the nature of competition for essential resources. However. Bradford. even though the reactor was operated under stable conditions. Horton. S. R (1980) Competitive exclusion. PC. ME. the model assumes a constant decay rate for all species. The results of our work suggests that the current activated sludge models (e. We speculate that this dynamic behavior could be caused. and phosphorusaccumulating organisms. Hugenholtz. ciliates (predators) and carnivores (top predators). References 1. Fredrickson. M (1971) Microbial Ecology.66 d. should also be considered. Wagner-Dobler. Burrell. Alexander. this study focused on the effects of competition within a single trophic level (bacteria). First. H. AG (1988) Limitation of growth rate by two complementary nutrients: some elementary but neglected considerations. In addition. OERTHER: IMPACT OF SRT ON 283 DIVERSITY 605 days. Finally. ASM1. Am Naturalist 115: 151–170 3. The model was only used to test the impact of a single operational parameter.g. Gwillian. in part. The results of the model simulation showed that for an SRT range of 2. Wat Sci Technol 37: 451–454 5. a number of limitations should be discussed.

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