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Municipal sewage wastewater treatment by activated sludge process: results

of a pilot scale study

Article · December 2019

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El Moussaoui & al./ Appl. J. Envir. Eng. Sci. 5 N°4(2019) 390-401

Municipal sewage wastewater treatment by activated sludge process:

results of a pilot scale study

T. El Moussaoui(a,b,c), M.O. Belloulid(a,b), Y. Jaouad(a,b) , L. Mandi(a,b) , N.

Ouazzani(a,b)

(a)
Laboratory of Hydrobiology, Ecotoxicology, Sanitation and Global Changes (LHEA URAC 33), Faculty of
Sciences Semlalia, University of Cadi Ayyad, Marrakech 40 000, Morocco
(b)
National Center for Studies and Research on Water and Energy (CNEREE), BP/511, Cadi Ayyad University,
Marrakesh 40 000, Morocco
(c)
School of Engineering & Innovation (E2IM), Private University of Marrakech (UPM), KM13, Route
d’Amizmiz , Marrakech 42 312, Morocco

* Corresponding author. E-mail address: [email protected] or [email protected]

Received 23 Sep 2019, Revised 13 Nov 2019, Accepted 23 Dec 2019

Abstract
The objective of the current research was to perform an experimental pilot study of municipal sewage
wastewater treatment by activated sludge process. To this end, the biomass growth, system
performance, flocs stealliabilityand microbial hygienic parameters were evaluated and assessed. The
results highlight an optimal growth of the biomass, which reached 4.83±0.25 gTSS.l-1 and 3.68±0.2
gVSS.l-1. The activated sludge performances in term of TSS, COD, BOD 5, TKN and TP were 92%,
94%, 97%, 85% and 16%, respectively. This value shows an efficient treatment of the wastewater
organic pollution and nutrients by activated sludge pilot system. In addition, the treated effluent
measured parameters concentrations were bellow Moroccan regulation for direct discharge into
environment. The sludge volume index (SVI) average concentrations were in the range of 80 mL.g-1 to
150 mL.g-1, indicating the good folcs settleability during the experimental period. Concerning
microbiological analysis, no significant reduction of all measured bacteria (p>0.05) was observed in
the treated wastewater. Therefore, the concentrations of all bacteria remained practically stable after
the treatment and were above Moroccan regulation guidelines for wastewater reuse in agriculture and
WHO guidelines for the safe use of wastewater. The results obtained from this pilot scale study
advocate that the activated sludge pilot system has performed adequately under conditions equal to
its original design criteria and complained with Moroccan applied discharge standards. On other hand,
to reuse treated wastewater in agriculture a disinfection process is recommended to ovoid public health
and environmental contamination.

Keywords: Municipal wastewater; Activated sludge; Performances; Stealliability; Microbiological

analysis.

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1. Introduction
The increase in water demand from population increase in arid and semi-arid zones makes converted
municipal wastewater a precious resource. The collection and treatment of municipal sewage and
wastewater is vital to public health and clean water. It is among the most important factors responsible
for the general level of good health in countries. Sanitation network collect sewage and wastewater
form homes, businesses, and industries and deliver it to wastewater treatment plant before it is
discharged to water bodies or land, or reused in irrigation of green spaces and/or agricultural purpose
[1-3].
Inadequate municipal sewage wastewater treatment leads especially to the degradation of conventional
water resources quality and environment in general. In many Mediterranean countries, such as
Morocco, rapid urbanization, industrial development and shortage of fresh water constitute a severe
problem. Ensuring water supplies has become an important issue for governments throughout the
Mediterranean region. However, global change will not only have impacts on the availability of water,
but also on water quality and ecosystem services [4]. The shortage of water has been affected by the
impact of climate change (increase of temperatures, variation of precipitations...). In the last fifty
years, the aggregate water demand has doubled resulting from demographic pressure and from the
development of water intensive activities such as tourism and some manufacturing in sectors such as
food, textiles and tanning. Indeed, most of the water is used in the agricultural sector, which presents
high rates of inefficiency [5].
In a local context, Morocco, as others Mediterranean countries is undergoing rapid socioeconomic and
technological changes that increase the pressure on its already structural water deficit [6]. The
availability, the protection and the management of water resources is a challenge in these regions. The
integrated water resources management and the adoption of emerging technologies for using fresh
and/or unconventional water resources more effectively is crucial. The increase in water consumption
and awareness of users to protect water resources and the environment is encouraging a greater
recovery of wastewater and more efficient and sustainable use of conventional water resources. The
treated urban wastewater reuse is an important part of the cycle and is a growing practice. The present
interest is due to social, economic and technical factors related to local conditions [7,8].
Wastewater treatment biotechnologies reduce organic load and suspended solids to limit pollution to
the natural environment. Several treatment technology includes physical, biological, and chemical
methods has been applied. Residual substances removed or created by treatment processes must be
dealt with and reused or disposed of in a safe way. Depending on its physicochemical and/or
microbiological characteristics, the purified wastewater is discharged to surface water or reused in
irrigation [9,10].Currently, the advancement of scientific knowledge and moral awareness has led to a
reduction of discharges through pollution prevention and recycling, with the noble goal of zero
discharge of pollutants.
The present work aims to perform an experimental study of municipal sewage wastewater treatment
by activated sludge process. Through the monitoring of biomass growth in the bioreactor of ASPP;
system performance; flocks stealliability and microbial hygienic parameters over the time. Further the
comparison of treated effluent physicochemical and microbiological quality parameters, regarding
national and international standards for discharge and/or discharge into environment.

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2. Materials and methods

2.1. Activated sludge pilot plant (ASPP)
The pilot plant activated sludge used in this experiment is presented in the Figure. 1

Figure 1. Schematic diagram of the activated sludge pilot plant ASPP

The pilot plant reproduces the operation of the activated sludge process full-scale plant. It consists of
an anoxic basin (18 l), an aerobic tank (60 l), a secondary sedimentation basin (30 l), a substrate vat
(100 l), a circulating pump and an aeration system (Figure.1). This installation aims to highlight the
main principles of biological treatment using activated sludge process.
The pilot is equipped with various analytical instruments, including probes for oxygen, pH and redox
potential. All equipment were connected to a central programmable logic controller (PLC) linked to a
supervisory control and data acquisition (SCADA): PLC SOFREL, which is a single program,
manages the automation and supervision providing a direct access and data storage.

2.2. Experimental setup

2.2.1 Activated sludge and wastewater source
For the start-up of the pilot plant of activated sludge, the aeration and the secondary sedimentation
basins were filled with activated sludge and wastewater collected from the aeration and secondary
sedimentation basins from Marrakech municipal activated sludge wastewater treatment plant. The
main physicochemical characteristics of the activated sludge and wastewater used in this experiment
are presented in table 1. The aeration basin of the pilot plant is filled with 60 L of the biomass (3.44
g.l-1) picked up from the aeration basin of the Marrakech wastewater treatment plant activated sludge.
The secondary sedimentation basin (30 l) is filed with wastewater collected from the secondary
sedimentation basin of the Marrakech wastewater treatment plant activated sludge.

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Table 1. Activated sludge and wastewater main physicochemical characteristics (average ± Standard
Deviation SD)

Influent WW Aeration tank Settling tank

-1
(g.l ) 0.222±0.03 3.5±0.5 -
-1
(g.l ) 0.158±0.01 2.85±0.3 -
-1
(mg.l ) 501.88±15 1545±102 30 ±1.5
-1
(mg.l ) 370±11 695±21 -
-1
(mg.l ) 76.53±5.5 39.25±3.1 11.63±2.01
-1
(mg.l ) 58.20±5.1 21.12±2.2 8.15±1.02
(mg.l-1) 5.12±0.5 10.62±1.1 9.52±0.8
-1
(mg.l ) 10.73±1 14.12±2.1 8.96±1.02
-1
(mg.l ) 9.53±0.7 11.87±1.4 7.63±1.1

2.2.2 ASPP preparation and monitoring

The operation of the activated sludge pilot system was started by inoculating activated sludge taken
from the aeration tank of the municipal activated sludge Wastewater Treatment Plant (WWTP) of
Marrakech (Morocco). The initial sludge total suspended solids (TSS) concentration was 3.45 g L -1 as
presented in Table.1
Wastewater after primarily decantation collected from Marrakech municipal WWTP was continuously
fed to the pilot plant using a peristaltic pump, whose physicochemical characteristics are presented in
Table.1
The two systems were in continuous operation in parallel for 4 months under the following operational
conditions:
 Recycling rate/flow rate were set during the experiment to 1, to have a standard behavior and
to ensure the presence of a significant amount of biomass. The reactor was at ambient
temperature during all the experiments approximately 25 °C.
 The hydraulic retention time (HRT) in the bioreactor was calculated using the working volume
divided total influent flow and recycling flow, it was about 10 hours.
 In the aeration tank, the oxygen was provided by an air bump regulated to maintain residual
dissolved oxygen concentration between 2 mgO2l-1and 3 mgO2 l-1.
 Treatment performance was monitored by following COD, BOD5, NH4, NTK and NO2, as
well as TN, pH values in the effluent three to five times a week.
 Nominal organic loading rate of 0.3 kgCOD.kgVSS-1.J-1has been maintained during the
experimental period, the substrate flow rate was regulateddepending on the influent COD
concentration.

2.3. Analytical methods

2.4.1 Physicochemical analysis
Samples were collected every working day from influent and effluent, as well as from biological tank,
to investigate biomass evolution and the system performances. Collected samples were stored at 4 °C

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in laboratory using plastic bottles for physicochemical analysis, and sterile glass bottles for
bacteriological studies.
Collected samples were analyzed according standards methods. The measured parameters were total
suspended solids ( ), volatile suspended solids ( ), chemical oxygen demand ( ), 5-day
biological oxygen demand ( ), total Kjeldahl nitrogen ( ),ammonium ( ), total
phosphorus ( , orthophosphates ( ), and sludge volume index ( [11,12,13].Temperature (T),
pH, electrical conductivity (EC) and dissolved oxygen (DO) were measured immediately via probes
installed in the ASPP.
2.4.3 Microbiological analysis
Bacteriological population analyzed were total count at 22°C and 37°C, fecal and total coliforms,
streptococci, intestinal enterococci, Escherichia coliand pathogens such as, Clostridium
perfringens,Staphylococcus aureus, Pseudomonas aerogenosaand Salmonella sp.
Bacteriological analysis methods were quantitative for the indicator organisms of faecal contamination
and qualitative for pathogens. The dilution method was used for the enumeration of microbial
population or the most probable number (MPN) technique for samples suspected to be highly
contaminated [14].
The analysis was done using the dilution method or the most probable number (MPN) technique for
the samples suspected to be highly contaminated [14]. Fecal coliforms, total coliforms, streptococci
and intestinal enterococci concentrations were measured by the MPN method. The others studied
bacteria concentrations were estimated by the dilution method. The removal of the microbiological
indicators was expressed as log units of colony forming units (CFU) per 100 mL (log CFU 100 mL−1).
The culture media used were as follows:
 Standard total Plate Count Agar PCA, for total bacterial count at 22°C and 37°C (Moroccan
Standard ISO 6222).
 Lactose TTC agar with Tergitol, for coliform counts (fecal and total coliforms), and soybean
casein digest agar for Escherichia coli (Moroccan Standard 08.0.124).
 Bileaesculin agar for group D streptococcicounts and Slanetz and Bartley medium for
enumeration of IE (Moroccan Standard 03.7.001).
 Baird Parker agar was used to select Staphylococusaureus. Identification was confirmed by
growing the strains on beef-heart infusion agar followed by a coagulase test with rabbit
plasma (Moroccan Standard ISO 6888-1-2008).
 Clostridium perfringenswas counted on sulphitepolymyxinsulphadiazine and cysteine agar
SPS agar. The mother solution was heat-treated at 80°C for 10 min (Moroccan Standard
08.0.125).
 Cetrimide agar was used to isolate Pseudomonas aerogenosa. After wards; serological
identification was made in a special medium King A (Moroccan Standard ISO 16266).
 Salmonella sp. strainswere isolated using Hektoen enteric Agar, after their enriching in the
selected Rappaport–Vassiliadis medium. Biochemical confirmation of isolated colony was
made using a Kligler Iron Agar in a slanted position “inoculated into the bottom of the tube”
(Moroccan Standard ISO 6579).

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2.4 Statistical analysis

The statistical significance of differences among means was determined by one-way analysis of
variance (ANOVA) followed by the Student–Newman–Keuls grouping test using SPSS (version 18.0),
the calculated means and standard deviations were also reported. All differences were considered
significant at 5%.

3. Results and discussion

3.1. TSS and VSS evolution in aeration tank of ASPP
In activated sludge process, the efficiency in the organic matter degradation and contaminants
removals in the activated sludge process depends on the amount of the biomass in the aeration basin.
However, the most employed parameter to quantify biomass amount in the bioreactor of an activated
sludge wastewater treatment plant is the amount of suspended solids, expressed as the total suspended
solids (TSS) and/or the volatile suspended solids (VSS).
The TSS and VSS analysis were performed every working day of the experimental campaign, in order
to assess and monitor the biomass growth in the aeration basin of the pilot system. In addition, the
VSS/TSS ratio was calculated.
Figure2. Shows the result of TSS and VSS and VSS/TSS ratio evolutions during the operation time of
this experiment. Throughout this experiment, the TSS and VSS average concentrations were ranged
from 4.83±0.25g.l-1to 2.88±0.12 g.l-1 and from 3.68±0.2 g.l-1to 2.29±0.1g.l-1, respectively. The average
VSS/TSS was about 72± 0.1%.
TSS g/l VSSg/l VSS/TSS
10 100%

8 80%
Concentration (g.l-1)

6 60% VSS/TSS (%)

4 40%

2 20%

0 0%
7 9 11 13 15 17 22 26 31
Time (Days)

Figure 2. Biomass evolution in aeration tank and VSS/TSS ratio over the time
The monitoring and assessment of TSS and VSS evolutions during this experiment report that is the
bioreactor of pilot system has performed adequately and at conditions equal to its nominal capacity
(design criteria), also the ratio VSS/TSS was optimal. This indicate the presence of an optimal amount
of biomass during this experiment. This result highlight a successful adaptation and selection of a
biomass able to remove effectively the organic load of municipal sewage wastewater.

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3.2 Performance assessment of the ASPP

3.2.1 COD and BOD5 abatement in the ASPP
Activated sludge refers to a mass of microorganisms cultivated in the treatment process to break down
organic matter into carbon dioxide, water, and other inorganic compounds. The COD and BOD 5are
used as the surrogate parameters to measure the organic matter available for the microorganisms. The
BOD5 test has been in use for more than a century, and consequently remains deeply entrenched in the
practice and experience of biological wastewater treatment. The COD analysis estimates the amount of
organic matter in wastewater in only three to four hours, rather than the five days required by the
BOD5 test, and can be used as an alternative.
COD results are typically higher than BOD 5 values, and the ratio between them will vary depending
on the characteristics of the wastewater. This ratio has been commonly used as an indicator for
biodegradation capacity [15].
Figure.3 depicts the BOD5 and COD temporal evolution and its removal efficiency in in the inflow
and outflow effluent in both experimental pilot systems during this experiment.
Influent Effluent Removal effeciency
700
100%
600
80%

Removal effeciency (%)

500
Concentration (g/l)

400 60%

300
40%
200
20%
100

0 0%
COD (mg/l) BOD5 (mg/l)

Figure 3. COD and BOD5 evolution and removal effeciency over time
The COD and BOD5 influent concentrations were 501.88±15 mg.l-1 and 370±11 mg.l-1, respectively.
In contrast, the output effluent concentrations were 30±1.5 mg.l -1 and 10.15±1.05 mg.l-1, in accordance
with the Moroccan applied discharges standards [9]. Moreover, the COD and BOD 5 removal
efficiencies were significant (p<0.05) 94% and 97%, respectively. This value shows an efficient
treatment of the wastewater organic pollution by conventional wastewater treatment plant
[8,15,16,17].

3.2.2 Nutrients evolution in the ASPP

3.2.2.1 Nitrogen evolution and abatement in the ASPP
Nitrogen removal has become an important part of wastewater treatment processes due to the
significant impact of nitrogen compounds ( , and ) on the aquatic environment and more

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stringent legislation on wastewater discharges. To meet this demand, the most commonly used method
for nitrogen removal is biological treatment based on aerobic nitrification and anoxic denitrification,
both of which may produce nitrous oxide (N2O) [15,18].
Figure 4 depicts TKN and NH4 contents evolution and abatementby the ASPP during the
experimental period.
The input average TKN and NH4 were 76.53 ± 5.5 mg.l-1 and 58.20 ± 5.1 mg.l-1 on average,
respectively. Whereas, the output effluent TKN and NH4 concentrations were on average 11.63 ± 2.01
mg.l-1 and 8.15 ± 1.02 mg.l-1, respectively. The TKN output concentration were bellow European
Standards (10 to 15 mg/L) [19] and Moroccan applied discharge standards (40 mg.L-1) [9].
The overall TKN and NH4 removal efficiencies were significant (p<0.05)85% and 86% respectively.
This indicates a good overall performance of the pilot plant activated sludge during this experiment
[15,16].

Influent Effluent Removal effeciency

90

80 80%

70

Removal effeciency (%)

Concentration (mg.l-1)

60 60%

50

40 40%

30

20 20%

10

0 0%
TKN NH4

Figure 4. Nitrogen (TKN and ) evolution and removal efficiency during the experiment

3.2.2.2 Phosphorus evolution and abatement in the ASPP

Controlling phosphorous discharged from municipal wastewater treatment plants is a key factor in
preventing eutrophication of surface waters resources. Phosphorous is one of the major nutrients
contributing in the increased eutrophication of lakes and natural waters. Its presence causes many
water quality problems and increased purification costs. Urban wastewater may contain from 5 to 20
mg.l-1 of total phosphorous.
The removal of phosphorous from wastewater involves the incorporation of phosphate into TSS and
the subsequent removal from these solids. Phosphorous can be incorporated into either biological
solids (activated sludge biomass) or chemical precipitates. In the biological removal of phosphorous,
the phosphorous in the influent wastewater is incorporated into cell biomass, which is subsequently
removed from the process as a result of sludge wasting. The reactor configuration provides the P
accumulating organisms (PAO) with a competitive advantage over other bacteria. In a conventional
activated sludge wastewater treatment plant, such as Marrakesh WWTP, phosphorus biological
removal performed mainly by microorganisms of activated sludge (PAO) [20,21,22].In activated

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sludge, bacteria only use enough phosphorus to satisfy their basic metabolic requirements, resulting in
typical removal rates of 20-40 % [23].
The average values of TP and PO43- concentrations evolution and abatement in the activated sludge
pilot plant ASPP are reported in the Figure 5.

Influent Effluent Removal effeciency

16
20%
14
Concentration (mg.l-1)

12

Removal effeciency (%)

16%

10
12%
8

6 8%

4
4%
2

0 0%
PT PO4

Figure 5. phosphorus (TP and ) evolution and removal efficiency during the experiment
The input effluent TP and average concentrations were 10.73±2.2 mg.L-1 and 9.53± 1.25 mg.L-1,
well the output effluent average concentrations were 8.96±1.01 mg.L -1 and 7.93 ± 1.35 mg.L-1,
respectively. The TP concentration in treated effluent is above the Moroccan applied discharge
standards (2 mg.L-1) [9]. In the bioreactor of ASPP, the TP and concentrations were about 14.12
-1 -1
mg.L and 11.81 mg.L more than the influent concentration because of storage capacity of
phosphorus and polyphosphate by the microbial biomass of activated sludge as reported in several
research studies [20-27].
The removal efficiencies of TP and by the activated sludge pilot plant ASPP were 16% and
20%, respectively. This result is comparable with those reported by Brett et al. (1997)[23].

3.3 Flocks stealliability

The extent to which activated sludge flocs settle within the secondary clarifiers directly influences the
concentration of suspended solids in the effluent and as a consequence the quality of the treated water
[28]. In return, producing a good effluent requires a manageable biological process and controlling the
reliable factors that affect the sludge biological properties and influence the characteristics of sludge, a
bad or good settling. As among the mainly performance-limiting factors, we denote toxicity, nutrients,
organic loading rate, temperature and mixed liquor volatile suspended solids concentration. Several
authors have reported the effect of a bad control of these limitations factors, in biomass properties
(break up of flocs or formation of unstable flocs particles) and normal flocs settling [29].
In the operation of activated sludge plants as well as in research studies, the Sludge Volume Index
(SVI) is used to characterize the settling properties of activated sludge. In the course of this
experiment, and in the order to assess the flocs stealliability sludge volume index (SVI) and diluted

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sludge volume index (SDVI) were performed as shown in Figure 5. The sludge volume index (SVI)
measured during this experiment highlighted well the good flocssettleability (80 mL.g-1 ≤ SVI ≤150
mL.g-1) [30].
200
SVI (mL g-1) DSVI (mL g-1)

150
Concentration (mL.g-1 )

100

50

0
7 8 9 10 11 12 13 14 15 19 20 21 23 27 29 31
Time (days)

Figure 5. Sludge volume index & diluted sludge volume index evolution during the experiment

3.4 Microbial hygienic parameters

The microbial analysis of influent and effluent wastewater from the activated sludge pilot plant is
showed in the Table.4.
Table 4. Concentrations of the microbiological parameters (Log colony forming units, CFU 100 mL −1)
(mean ± standard deviations)
RWW (input) TWW (effluent)
Total coliforms 5.87 ± 0.15 5.19 ± 0.11
Fecal coliforms 4.95 ± 0.05 4.28 ± 0.02
Streptococci D 4,58 ± 0.12 3.93 ± 0.03
Clostridium perfringens 4.61 ± 0.18 5.09 ± 0.01
Pseudomonas aeruginosa 4.70 ± 0.18 5.10 ± 0.10
Total bacterial count (37°C) 6.68 ± 0.50 5.55 ± 0.40
Total bacterial count (22°C) 6.11 ± 0.12 5.24 ± 0.24
Escherichia coli 4.64 ± 0.04 4.35 ± 0.05
Intestinal enterococci 4.07 ± 0.41 3.59 ± 0.59
Staphylococcus aureus 4.62 ± 0.04 3.89 ± 0.02
Salmonella spp. + +
RWW: Raw wastewater
TWW: Treated wastewater

The average concentrations of measured bacterial indicator and pathogens in raw wastewater were
5.87± 0.15 log units for Total coliforms, 4.95±0.05 log units fecal coliforms, 4.64±0.04 Escherichia

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coli, 6.68±0.50 log units and 6.11±0.12 log units for total bacterial count 37°C and 22°C respectively,
4.78±0.12 log units for Streptococci D, 4.61±0.18 log units for Clostridium perfringens, 4.62±0.04
long units for Staphylococcus aureus.Also the Salmonella spp, determined by qualitative methods,
was detected in raw wastewater and even in treated water
No significant reduction of all measured bacteria (p>0.05) was observed in the treated wastewater
(effluent). Therefore, the concentrations of all bacteria remained practically stable after the treatment;
this result is expected because we did not apply a tertiary treatment in our case. However, the input
concentrations of indicators bacteria are above Moroccan regulation guidelines for wastewater reuse in
agriculture [31] and WHO guidelines for the safe use of wastewater [10].

4. Conclusion
The results reported in this experimental pilot scale study of municipal sewage wastewater treatment
by activated sludge process highlight an optimal biomass growth, high abatement of organic matter
and optimum flocs seatliability. Indeed, The ASPP performances in term of TSS, COD, BOD 5, TKN
and TP were 92%, 94%, 97%, 85% and 16%, respectively. In addition, the sludge volume index (SVI)
average concentrations were in the range of 80 mL.g-1 to 150 mL.g-1indicating good folcs settleability.
Microbiological analysis of raw and treated raw wastewater showed no significant reduction of all
measured bacteria (p>0.05) after the treatment. Therefore, the concentrations of all bacteria remained
practically stable after the treatment and were above Moroccan regulation guidelines for wastewater
reuse in agriculture and WHO guidelines for the safe use of wastewater. To this end, a disinfection
process must be applied before reusing treated municipal sewage wastewater in agriculture.
Acknowledgements
The authors would like to knowledge the National Center for Studies and Research on Water and
Energy (CNEREE) Cadi Ayyad University and the FAO project GCP/RAB/013/ITA for the financial
and the technical support.
Declarations of interest
The author declares that there is no conflict of interest whatsoever.

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