Determining the causes of the deterioration of granules in an

aerobic granular sludge continuous flow system

Alallana E.*, Franchi O.**, Pavissich J.P. **, Crutchik D.**, Da Silva C.*, Guerrero L.*, Pedrouso A.***,
Val del Río A.***, Mosquera-Corral A.*** and Campos J.L.**

*Chemical and Environmental Engineering Department, Technical University Federico Santa María, Chile, Ave.
España 1680, Valparaíso, Chile (esteban.alallana.14@sansano.usm.cl; cristopher.dasilva@usm.cl;
lorna.guerrero@usm.cl)
**Facultad de Ingeniería y Ciencias, Universidad Adolfo Ibáñez, Avda. Padre Hurtado 750, Viña del Mar, Chile
(oscar.franchi@edu.uai.cl; juan.pavissich@uai.cl; dafne.crutchik@uai.cl; jluis.campos@uai.cl)
***Department of Chemical Engineering, Institute of Technology, Universidade de Santiago de Compostela, E-
15705 Santiago de Compostela, Spain (alba.pedrouso@usc.es; mangeles.val@usc.es; anuska.mosquera@usc.es)

Abstract
The application of the aerobic granular sludge technology in continuous flow systems is still a
challenge. In this work, a continuous reactor of 25 L, with six baffles to promote a feast/famine
regime, was inoculated with mature granules. When the upflow velocity of 2.9 m/h was imposed
in the settler, the flocculent biomass ended up predominating in the system while, when this
upflow velocity was increased up to 6.0 m/h, granular biomass maintained its predominance, but
its sludge volume index (SVI) worsened along the operation time. This fact could be related to the
low substrate concentration present during the feast period. A model to explain the competition
between the granular and flocculent biomass was developed. According to this model, the ratio
between the solids retention time of granular and flocculent biomass, obtained in each experiment,
defines the predominant type of biomass inside the reactor.

Keywords
Continuous flow; feast/HRT ratio; granular biomass; solids retention time; substrate concentration.

INTRODUCTION
The constant increase of the urban population brings with it the need to increase the capacity of the
existing wastewater treatment plants (WWTPs). Since, in large cities, the available area to expand
the WWTPs is limited, the most suitable alternative to increase their treatment capacity seems to be
the change of the conventional activated sludge systems (CAS) by other higher-rate technologies. In
this context, a successful alternative technology developed during the last two decades corresponds
to the aerobic granular sludge (AGS) (Hamza et al., 2022). AGS offers improved settling properties
in comparison to CAS, which allow increasing the suspended solids concentration in aeration tanks
and, therefore, increasing the treatment capacity of the system. Although specific conditions to
promote aerobic granule formation are well known in sequencing batch reactors (SBR), the
conditions to achieve a rapid and stable granulation in continuous flow systems are still under
investigation and have been the focus of many AGS studies (Xu et al., 2022). On basis of these
observations, the aim of the present work is to define the operating conditions of a continuous flow
system that allow maintaining the physical stability of aerobic granules such as in an SBR.

MATERIALS AND METHODS

A continuous reactor of 25 L with six baffles was operated to determine the suitable operational
conditions to maintain the long-term stability of granular biomass. Granular biomass taken from a
25 L SBR reactor operated in parallel to the continuous flow reactor was used as inoculum. Both
systems were fed with a synthetic medium simulating the composition of domestic wastewater
(Jungles et al., 2011). The operating conditions imposed in the continuous flow reactor and the
inoculum characteristics of the 3 experiments carried out are detailed in Table 1.
Table 1. Operating conditions and inoculum characteristics of the experiments.
Experiment Operating conditions Inoculum
Flow rate Upflow velocity in HRT Initial concentration SVI
(L/d) the settler (m/h) (h) (TSS/L) (mL/g TSS)
1 100 2.9 6.0 3.2 26
2 200 6.0 3.0 1.9 56
3 200 6.0 3.5* 1.5 60
*An anaerobic chamber of 4 L was added to the system before the aerobic reactor.

RESULTS
In all the experiments, the system was able to remove more than 95% of the organic matter while
nitrogen removal efficiency was around 34% and was due to assimilation, since neither nitrate nor
nitrite was detected. None of the operating conditions tested were suitable to maintain the physical
stability of the granules for a period longer than 22 days. In the first experiment, the hydraulic
selection pressure imposed in the settler was not able to promote the wash-out of the flocculent
biomass. Also, since flocs grow faster than granular biomass, its overgrowth probably limited the
availability of substrate for granular biomass development. In the case of the second and third
experiments, the hydraulic selection pressure imposed was increased and flocculent biomass was
successfully washed-out. This promoted the growth of aerobic granules (Figure 1). In the second
experiment the average diameter of granular biomass increased from 0.5 up to more than 2.0 mm.
Nevertheless, their appearance was fluffy, with an SVI of 109 mL/g TSS. A similar behavior was
observed during the third experiment but, in this case, the average diameter and the SVI were lower
(1.80 mm and 82 mL/g TSS, respectively) than in the previous experiment. This difference could be
attributed to the implementation of an anerobic contact tank during the third experiment.
A B C

D E F

Figure 1. Aspect of biomass. Inoculum: A) Experiment 1; B) Experiment 2; C) Experiment 3. End of the

experiment: D) Experiment 1; E) Experiment 2; F) Experiment 3.
In addition to the hydraulic selection pressure, maintaining a suitable feast/famine regime by means
of the hydraulic retention time (HRT) is a key operational condition to promote the formation of
stable granular biomass. In order to determine if the values of the feast/HRT ratio during the
different experiments were suitable to favor the growth of granular biomass over the flocculent one,
a conceptual model was developed based on the growth rate of both kinds of biomass. This model
allowed calculating the solids retention time (SRT) required by each type of biomass to grow under
certain feast/HRT value. The ratio between the SRT values calculated for granular and flocculent
biomass (SRTg/SRTf) would define the conditions in which the granular biomass would begin to
predominate over flocs for a certain value of the feast/HRT ratio (Figure 2). From the COD
concentration profile measured along the reactor, a feast/HRT value of 0.14 was determined during
the first experiment and of 0.29 during the second and third experiments. Considering the SRT
calculated for each type of biomass during the experiments, the model results suggest that the
selective hydraulic pressure imposed during the first experiment was not strict enough to achieve
the predominance of the granular biomass, while the up-flow velocity imposed in the settler in both
experiments 2 and 3 allowed a suitable uncoupling of the SRT of granular and flocculent biomass
(Figure 2). However, even though the operating conditions of the two last experiments seemed
suitable for the development of the granular biomass, the physical characteristics of the granules
generated were not the desired ones (i.e. high SVI, long-term instability). This suggests that the
selective hydraulic pressure and the feast/famine regime were suitable for granulation, but not
enough to generate granular biomass with good physical characteristics.
30

25

20
SRTg/SRTf

15

10

0
0.0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1.0
Feast/HRT

Figure 2. Minimum solid retention time of granular and flocculent biomass ratio required for the
predominance of the granular biomass (dashed line); solid retention time of granular and
flocculent biomass ratio obtained in: Experiment 1 (•); Experiment 2 (); and Experiment 3 ().
Since the deterioration of the physical properties of granules observed during experiments 2 and 3
could be related with the growth of ordinary heterotrophic biomass (XH) over accumulating
heterotrophic biomass (XSTO), a simulation based on a diffusion-reaction model was carried to
determine how the substrate is consumed inside the granule. The simulation done under the feast
phase conditions of both experiments (65 mg COD/L and HRT of 20 minutes) shows that the
generation of XH is 1.46 times higher than that of XSTO and, therefore, the granulation process
would not be favored. In order to determine the influence of the substrate concentration on the
granulation process, a new simulation was performed including the granule size range observed in
the experiments (Figure 3). This simulation showed that for a granule diameter of 1.7 and of 3.1
mm substrate concentrations lower than 100 mg COD/L promote the growth of the ordinary
heterotrophic biomass over the accumulating heterotrophic biomass (XH/XSTO higher than 1). Also,
for these given sizes the XH/XSTO is lower than 1 at substrate concentrations higher than 250 mg
COD/L. At higher concentrations the substrate gradient may not limit the growth of accumulating
heterotrophs inside the granules. Thus, the configuration of a continuous flow reactor used to
generate granular biomass not only should have a suitable feast/famine regime, but also maintain an
adequate substrate concentration during the feast period in order to promote the growth of the
accumulating heterotrophic biomass.
 3.5
3.0
XH/XSTO [-] 2.5
2.0
1.5
1.0
0.5
0.0
0 50 100 150 200 250 300 350 400 450 500
COD [mg/L]
Figure 3. Influence of substrate concentration on the biomass concentration ratio between ordinary
heterotrophs and accumulating heterotrophs (granule diameter: 3.1 mm (continuous line); 1.7 mm
(dashed line); 0.5 mm (dotted line)).

ACKNOWLEDGEMENTS
This research was funded by the Chilean Government through the Projects ANID/FONDECYT/1200850 and CRHIAM
Centre grant number ANID/FONDAP/15130015, and by the European Commission LIFE ZERO WASTE WATER
[LIFE19ENV/ES/000631] project. The authors from Universidade de Santiago de Compostela belong to the Galician
Competitive Research Group [GRC ED431C-2021/37]. Esteban Alallana would like to acknowledge Postgraduate and
Programs Department of Technical University Federico Santa María for their support through a PIIC grant. Alba
Pedrouso would also like to acknowledge the Xunta de Galicia for their support through a Postdoctoral Fellowship
(ED481B-2021-041).

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