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Bioaugmentation of a sequencing batch reactor with Archaea for the treatment

of reject water

Article  in  Journal of Water Chemistry and Technology · July 2016

DOI: 10.3103/S1063455X16040093

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ISSN 1063455X, Journal of Water Chemistry and Technology, 2016, Vol. 38, No. 4, pp. 238–243. © Allerton Press, Inc., 2016.

BIOLOGICAL METHODS
OF WATER TREATMENT

Bioaugmentation of a Sequencing Batch Reactor with Archaea

for the Treatment of Reject Water1
A. Szajaa, G. Lagóda, *, J. Drewnowskib, F. Sabbac
aLublin University of Technology, Poland
bGdansk University of Technology, Poland
c
University of Notre Dame, Department of Civil and Environmental Engineering and Earth Sciences, USA
*email: g.lagod@pollub.pl
Recieved December 23, 2015

Abstract—In this study, the bioaugmentation of a sequencing batch reactor (SBR) for the treatment of
reject water from wastewater treatment plant was evaluated. For the bioaugmentation step a product con
taining an enrichment of microorganisms from the Archaea domain was used to enhance the performance
of the reactor for treating reject water. The experiment was carried out in two parallel labscale sequencing
batch reactors. The first one (SBR A) was bioaugmented with a suspension of microorganisms from the
Archaea domain, while the second reactor (SBR B) was not bioaugmented. The results here presented
show that the SBR technology could sustain efficient NH4+–N and chemical oxyden demand removal
rates and can be applied for the treatment of reject water. Moreover, the addition of microorganisms
belonging to the Archaea domain improved the SBR overall operation, especially when the loading in the
influent was increased. Administering Archaea to the reactor had also a positive effect on ammonia oxida
tion as well as on the nitrite removal.
DOI: 10.3103/S1063455X16040093
Keywords: reject water, bioaugmentation, sequencing batch reactor wastewater treatment plant, Archaea.

INTRODUCTION
Over the last few decades, wastewater treatment strategies have mostly focused on the process efficiency.
Additionally, with the improvement of the effluent quality the amount of wastewater treatment byproducts,
i.e. sewage sludge, have also significantly increased [1, 2]. Generally, the sewage sludge treatment includes
physicalchemical and biological methods, where the solid phase is separated from water. The most common
applied processes are: thickening, anaerobic digestion, chemical conditioning, and dewatering as well as dry
ing. During sludge processing the highstrength wastewater, called reject water, is generated [3]. This water is
characterized by high concentration of ammonium ranging from 750 to 1500 mg/L and phosphorus up to
130 mg/L [4–6]. Moreover, the variability of the composition and the irregularity in the flow of the streams
represent a challenge for a wastewater treatment plant (WWTP) performance [7]. Although the flow of reject
water often might be relatively small (around 2% of total flow) this type of wastewater could have a significant
influence on the overall wastewater treatment plants operation. Numerous issues including process upset,
overloading, and an increase in operational and maintenance costs [8, 9] might arise when treating this type
of waste. Commonly, the reject water is returned to the influent of the WWTP without any pretreatment, where
the possibility the application of conventional activated sludge system becomes relatively limited. This system
is mainly applied for treating wastewaters with low nitrogen concentrations (less than 100 mgN/L) and tem
peratures oscillating about 15°C [10, 11]. Therefore, a separated treatment for reject water draws more atten
tion. Different solutions have been proposed; these include physicalchemical methods, e.g. ammonia strip
ping [12] and struvite precipitation [13]. However, when these methods are compared with biological technol
ogies, they tend to be relatively expensive [8]. For nitrogen removal biological sidestream processes have also
been developed. These processes include anaerobic ammonium oxidation (ANAMMOX) [14], single reactor
system for high activity ammonium removal over nitrite (SHARON) [15], SHARON process combined with
ANAMMOX process [16] as well as completely autotrophic nitrogen removal over nitrite (CANON) and oxy
gen limited autotrophic nitrificationdenitrification (OLAND) technology [17, 18]. These technologies could
be operated in continuous and sequencing batch reactor (SBR) mode. The latter is based on the activated
sludge system, operated through a series of cycles that occur within the same tank. Because of lower costs as
1
The text was submitted by the authors in English.

238
 BIOAUGMENTATION OF A SEQUENCING BATCH 239

well as a greater operational flexibility than conventional plugflow biological treatment methods, this tech
nology has been widely applied in wastewater treatment [19–22]. These advantages, including the possibility
of handling shifts in both hydraulic and organic loadings, make the SBR an ideal candidate for the treatment
of reject water. Moreover, the application of bioaugmentation in sidestream processing, i.e. Bioaugmentation
Batch Enhanced (BABE) technology, has been reported [23]. The bioaugmentation technique is known as an
advanced method that introduces specific strains or mixed cultures into a biological system with the goal of
increasing its activity and performance [24–26]. Furthermore, this technique has been widely applied to aer
obic systems to improve process stability and enhance removal efficiency but also solve some practical prob
lems such as flow fluctuations, shock loads of specific contaminants or toxic exposure [27, 28].
In this study the bioaugmentation of a SBR for the treatment of reject water was proposed. For the bioaug
mentation step the product Arkea® was chosen and supplied to the system. The product consisted of micro
organisms from the Archaea domain; these microorganisms have been reported in literature to play an impor
tant role in the ammonia removal from wastewater [29–33].

EXPERIMENTAL

Wastewater and Seeding Sludge

Reject water, wastewater and seed sludge samples were obtained from the municipal WWTP Hajdów
located in Lublin (Poland). This WWTP applies primary and secondary treatment and is a mechanicalbio
logical plant that employs a modified Bardenpho method [34]. The plant treats a daily wastewater volume of
about 65000 m3. The wastewater used in the experiment was sampled from the effluent of the primary clarifier.
The reject water was collected from the dewatering belt press that treated sludge from mesophilic anaerobic
digesters (35°C). Both samples were collected twice a week, labeled and stored under refrigeration at 4°C. The
detailed composition of these samples is shown in the table. Both SBR were seeded with activated sludge from
the aeration tank operating at a temperature around 20°C [35]. The seeding sludge was characterized by mixed
liquor suspended solids (MLSS = 3.19 g/L) and mixed liquor volatile suspended solids (MLVSS = 2.43 g/L),
the food to microorganism ratio (F/M ratio = 0.11 gBOD5 /gMLVSS·d), sludge volume index (SVI =
234 ml/L) and a sludge retention time (SRT) of 14.9 d. For the bioaugmentation step the commercial product
Arkea® was used and prepared as liquor fed in continuous mode. A 250 ml aliquot of this product was used in
the experiment. The prepared suspension of bioaugmentation product was characterized by the following
parameters: COD = 22 mg/L, pH 7.16, TS = 0.47 g/kg and VS = 0.042 g/kg.

Characteristics of the wastewater used in the experiment (the mean value and standard deviation are given)
NO3––N NO2––N NH4+–N
Sample COD pH TSS, mg/L Turbidity(NTU)
mg/L
Reject water 110.20 ± 33.72 0.71 ± 0.23 0.22 ± 0.09 214.13 ± 17.00 8.06 ± 0.14 18.00 ± 8.76 13.15 ± 4.39
Wastewater 624.33 ± 34.54 0.69 ± 0.14 0.22 ± 0.004 59.89 ± 9.40 7.83 ± 0.07 315.17 ± 22.16 164.17 ± 16.75

Analytical Methods
During the experiment the following parameters were measured: COD, NO3––N, NO2––N, NH4+–N.
These analyses were performed for each SBR cycle by means of a spectrophotometer (Hach Lange UVVis
DR 5000) using Hach standard cuvette tests. The MLSS and MLVSS were determined according to Polish
standard methods. Activated sludge samples were analyzed once a week. Dissolved oxygen (DO) as well as pH
were constantly monitored by a LDO probe (HQ 40D HachLange). For turbidity a Turbidimeter CyberScan
TN 100 was used.

Experimental Reactors and SBR Operation

In the experiment two parallel labscale SBR were used. Each of them had a working volume of 8 L, and a
total volume of 10 L, with a diameter of 0.22 and height of 0.35 m. The reactors were equipped with a mechan
ical stirrer, air membrane diffuser and a heating/cooling device (water bath with controlled and regulated
temperature) (Fig. 1).

JOURNAL OF WATER CHEMISTRY AND TECHNOLOGY Vol. 38 No. 4 2016

240 SZAJA et al.

1
2

3
4

Fig. 1. Scheme of the labscale sequencing batch reactors: 1—electric motor driving the mixing system, 2—distribution tubes
for pressured air, 3—bioraction chamber, 4—water bath with controlled and regulated temperature, 5—lowspeed blade stir
rer, 6—membrane diffuser, 7—membrane supercharger supplying aeration system with pressured air.

The reactors were operated with 12 h cycles that were divided into the following distinct phases: fill
(30 min), react (stirring + aeration, 120 + 420 min), settle (90 min), decant (30 min) and discharge
(30 min). In both chambers, during the experiment the temperature was kept constant at 20 ± 0.1°C and
pH was monitored (mean value of pH 8.05 ± 0.2). DO concentration was maintained at range about
2 mg O2/L. The two SBR were operated under the same conditions, however SBR A was bioaugumented.
The reactor SBR A was fed with 2.5 L of presettled wastewater, 0.4 L of reject water and 0.25 L of sus
pension containing the Archaea used for bioaugmentation. The second reactor, SBR B, was fed with the
same wastewater and reject water volume, and the bioaugmentation product was in this case replaced
with an equal amount of distilled water. The activated sludge used in this experiment was acclimated with
the bioaugmentation product as well as the reject water for about 20 days. During the startup of the reac
tors 250 mL of the reject water was added to both SBR. After this phase, both SBR were operated for 55
days. The hydraulic retention time (HRT) of both reactors was kept constant at 3 days.

RESULTS AND DISCUSSIONS

The experimental data of the two SBR are summarized in Fig. 2 to 4. As shown in Fig. 2, the bioaugmented
SBR (SBR A) effluent average ammonium concentration was 2.49 ± 2.13 mg/L. In the case of the nonbio
augmented reactor (SBR B) a significant reduction of this parameter was also observed, with an average value
of 4.33 ± 3.83 mg/L. The ammonium removal efficiency was 99.1 and 98.4%, in SBR A and SBR B—respec
tively. The effective elimination of this parameter occurred mainly by microbial nitrification to nitrate [3]. A
similar reduction of this parameter (more than 95%) was observed for other technologies applied for the treat
ment of high ammonium wastewater as well [20, 23]. Additionally, it is worth mentioning that there is a pos
sibility of existence in the bioreactors of WWTP Hajdów ANAMMOXprocesses, the comparable trend was
observed in the research presented by Supura and Demchina [36]. It is noteworthy that the bioaugmented
reactor was able to withstand and tolerate an influent loading shock that occurred between day 7 and 14 (see
Fig. 2).
A similar trend was also observed for COD in the SBR effluent. Its value varied from 19.5 to 74.3 mg/L
for the bioaugmented reactor and from 10.4 to 71.7 mg/L for the nonbioaugmented one. The COD
removal was slightly higher in SBR B (85.24%) than in SBR A (84.57%). Moreover, the ammonium con
centration, as well as COD, for the reject water (see table) was lower than what previously found from
other researchers [4, 37–39]. However, the share of this sidestream was significant and accounted for
8.0% of the total influent flow in the Hajdów WWTP, and in some cases it reached as high as 10%. Addi
tionally, the presletted had high ammonium content. The high content in ammonium was due to the
setup of the wastewater plant that did not use any pretreatment system for the reject water and directed
the whole stream at the beginning of main technological line of wastewater purification.

JOURNAL OF WATER CHEMISTRY AND TECHNOLOGY Vol. 38 No. 4 2016

 BIOAUGMENTATION OF A SEQUENCING BATCH 241

25.0

Concentration [mg/L]
15.0

5.0

0 10 20 30 40 50
Time, [d]
bioaug. NO3  N 
bioaug NO2  N bioaug. NH4  N
+
  +
nonbioaug. NO3  N nonbioaug. NO2  N nonbioaug. NH4  N

Fig. 2. Nitrogen species concentration for the effluent of SBR A (bioaugmented) and SBR B (nonbioaugmented).

80.0

60.0
Concentration [mg/L]

40.0

20.0

0
0 10 20 30 40 50
Time [d]
bioaug. COD bioaug. TSS nonbioaug. COD nonbioaug. TSS

Fig. 3. Concentrations of chemical oxygen demand and total suspended solids in the effluent of the SBR A (bioaugmented)
and SBR B (nonbioaugmented).

4.0
8.0

3.0
Turbidity [NTU]

6.0
pH

2.0
4.0

2.0 1.0

0 0
1 6 11 16 21 26 31 36 41 46 51
Time [d]
bioaug. pH nonbioaug. turbidity nonbioaug. pH bioaug. turbidity

Fig. 4. The turbidity and pH in the effluent of the SBR A (bioaugmented) and SBR B (nonbioaugmented).

The average nitrate concentration was 20.78 ± 2.88 mg/L in SBR A and 19.79 ± 3.81 mg/L in SBR B,
respectively. In the bioaugmented SBR a lower influent NO2––N concentration (0.27 ± 0.08 mg/L) was
obtained. For SBR B the average concentration was 0.37 ± 0.12 mg/L. Significant removal efficiency was also

JOURNAL OF WATER CHEMISTRY AND TECHNOLOGY Vol. 38 No. 4 2016

242 SZAJA et al.

observed for the TSS with a final value of 91.2% (4.77 ± 1.7 mg/L) for SBR A and 92.1% (4.27 ± 2.26 mg/L)
for SBR B. pH was quite stable for both SBR. Its value ranged between 7.77 and 8.25 for SBR A and between
7.85 and 8.28 for SBR B.
Additionally, the turbidity of the wastewater effluent was monitored during the experiment (see Fig. 3).
SBR A and B behaved differently with regards to this parameter. For the bioaugmented reactor the turbidity
was 2.22 ± 0.61 Nephelometric Turbidity Units (NTU) and 1.96 ± 0.53 NTU for the nonbioaugmented one.
Finally, SBR A achieved more stable conditions as well as it was more resistant to adverse conditions (i.e.
increasing N loading) than SBR B. In case of all analyzed parameters, it was noticed that the coefficient of
variation and standard error were higher in case of nonbioaugmented reactor (SBR B), which may indicate
greater stability process for the bioaugmented reactor (SBR A).
The increasing ammonium loading in the influent of SBR did not affect the SBR operation. The reduction
of the measured parameters showed a similar trend of the SBR that treated municipal wastewater without
reject water addition (study where identical experimental conditions were used: HRT, temperature, pH,
MLVSS etc.) [40, 41].

CONCLUSIONS
In this study, the bioaugmentation of SBR with Archaea for the treatment of reject water was investigated.
The following conclusions can be drawn:
– findings from this study showed that the SBR technology could efficiently remove NH4+–N, COD and
TSS parameters from wastewater generated during sludge processing;
– the bioaugmentation with Archaea had an influence on the reject water treatment. This step improved the
SBR operation, especially in case of an increase of the loading in the influent. It also had a positive effect on the
ammonia oxidation and nitrite removal, showing overall higher removal efficiency;
– the proposed system could be adapted to existing wastewater treatment plants, which have strict effluent
nitrogen requirements or exposed to N loading fluctuations. It doesn’t require any modification in construc
tion of conventional SBR;
– this technology could be applied to systems treating wastewater rich in reject water and for wastes that
are recirculated to the main stream of the WWTP as well as to separate treatment of this sidestream;
– although this is a preliminary study, further research evaluating potential impacts of increasing flows of
reject water is needed.

ACKNOWLEDGMENTS
The authors would like to thank Agnieszka Montusiewicz for providing the Arkea® product. This project
was financed with the support from the National Centre of Science (Poland), No. 7405/B/T02/2011/40.

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