Journal of Environmental Management 269 (2020) 110714

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Journal of Environmental Management

journal homepage: http://www.elsevier.com/locate/jenvman

Research article

Full-scale sludge reduction in the water line of municipal wastewater

treatment plant
Claudio Di Iaconi *, Marco De Sanctis , Valerio Guido Altieri
Water Research Institute, C.N.R, Viale F. De Blasio 5, 70123, Bari, Italy

A R T I C L E I N F O A B S T R A C T

Keywords: Nowadays, sludge management represents one of the most critical challenges in the field of sewage treatment for
Full scale application economic and environmental impacts. Therefore, the reduction of sludge has become a major issue for the op­
Granular sludge system erators of municipal wastewater treatment plants. In the present paper, a new system, whose acronym is MULESL
Maintenance metabolism
(MUch LEss SLudge), is proposed and tested at full scale for reducing the quantity of sludge in the water line of
Sewage sludge
the sewage treatment plant. MULESL system takes the advantage of maintenance metabolism to significantly
Sludge minimization
reduce the sludge production. The effectiveness of MULESL system in removing the typical pollutants and
reducing sludge production was evaluated at full scale by using 3500 PE unit located in Putignano’s WWTP
(Puglia, Italy). This unit was obtained by retrofitting an existing activated sludge basin. The results obtained over
1-year period, during which MULESL unit treated the effluent of the preliminary treatment step, have indicated
that it was characterized by a specific sludge production as low as 0.13 kg of dry sludge per kg of COD removed;
77% lower than that recorded for primary and secondary treatments of the conventional plant during the same
period. This sludge reduction was obtained with a plant volume 27% smaller than that of the conventional water
line. Furthermore, the organic matter of the sludge was already stabilized, thus allowing to save investment costs
for digestion process facilities. Finally, MULESL unit guaranteed a mean removal efficiency higher than 95% for
COD, BOD5, TSS, TKN, NH3 and TN.

1. Introduction
The management of sludge originating from wastewater treatment
plants (WWTPs) is currently one of the most critical issues of the whole
treatment cycle of municipal sewage. In fact, although the sludge vol­
ume produced by wastewater treatment plants represents only 1% of the
volume of sewage flowing in the plants, its treatment and final disposal
entail very high capital and operating costs (up to 60% of total treatment
costs; Wei et al., 2003; Andreoli et al., 2007). Unfortunately, these fig­
ures are expected to increase further, as a result of progressively more
stringent effluent and sludge regulation criteria together with the
growing number of WWTPs (Collivignarelli et al., 2019). Therefore,
among sewage treatment plants managers there is currently a strong
interest in technological solutions to reduce the production of sludge to
be disposed of.
Several approaches for sludge minimization have been proposed in
the last decade although their application at the full scale is still rela­
tively limited. These approaches can be classified on the basis of the
location of the plant where minimization takes place. Based on this
criterion, the current available approaches are divided in two great
groups: those acting in the water line of sewage treatment plant and
those acting instead in the sludge line (Wang et al., 2017).
Usually, they are not implemented simultaneously in the same
WWTP. The first ones, however, are particularly interesting since they
handle the problem of the sludge at its origin, thus reducing the quantity
to send to the sludge line of the plant.
Most of the proposed techniques for reducing sludge production in
the water line are based on mechanisms such as cell lysis-cryptic growth
and maintenance metabolism (Foladori et al., 2010).
The cryptic growth mechanism exploits the reutilization of intra­
cellular material released during cell lysis (Hao et al., 2010). The tech­
nologies based on cell lysis-cryptic growth are implemented in the
return activated sludge (or a fraction of it) of the biological reactor. The
lysed sludge is then recycled to the main biological reactor where the
reutilization of the organic material released during bacteria lysis oc­
curs. A portion of this material is then oxidized in the catabolism phase,
which leads to the reduction of sludge production.
The technologies based on this mechanism are distinguished by the

* Corresponding author.
E-mail address: claudio.diiaconi@ba.irsa.cnr.it (C. Di Iaconi).

https://doi.org/10.1016/j.jenvman.2020.110714
Received 5 February 2020; Received in revised form 28 April 2020; Accepted 4 May 2020
Available online 27 May 2020
0301-4797/© 2020 Elsevier Ltd. All rights reserved.
C. Di Iaconi et al. Journal of Environmental Management 269 (2020) 110714

type of treatment that promotes the lysis that can be a chemical (such as 2. Materials and methods
ozonation, chlorination, Fenton treatment, etc; Saby et al., 2002; Sievers
et al., 2004; Gardoni et al., 2011), physical (ultrasound, thermal, elec­ 2.1. Plant description and operation
trical, etc; Pilli et al., 2011) or biological (i.e., anaerobic side-stream
reactor; Romero et al., 2015; Ferrentino et al., 2016) method. Howev­ In 2016 the first MULESL demonstrative unit with a treatment ca­
er, only a few of them have been applied so far at full scale as the pacity of 3500 person equivalents (PE) was built by retrofitting an
chemicals and operating conditions necessary for achieving significant existing activated sludge basin (670 m3 volume) of Putignano’s WWTP,
reductions of sludge production can negatively affect the purification located in sourthern Italy.
process as well as make the process expensive (Foladori et al., 2010; The water line of the existing Putignano’s WWTP (28,000 PE) con­
Wang et al., 2017). sists of preliminary treatment stage (for screening, desanding and
The processes that take advantage of the maintenance metabolism equalization operations), primary sedimentation (single clarifier with
are based on the increase of biomass concentration in the biological 472 m3 volume), biological stage with anoxic (1270 m3 volume) and
basin (that means long sludge retention times, SRT) in order to reduce aerobic (4570 m3 volume) compartments for carbon and nitrogen
the net growth yield. Maintenance energy mechanism is based on the removal, based on activated sludge process, secondary sedimentation (2
fact that microorganisms prefer to satisfy the energy requirement for clarifiers, total volume of 924 m3) and tertiary treatment, for phos­
non-growth activities before producing new cells (Wei et al., 2003). In phorous removal and disinfection. The sludge line is based on anaerobic
fact, it is well know that decreasing substrate/biomass ratio it is possible digestion (2 digesters, total volume of 2610 m3) and centrifuge dew­
to reduce the observed growth yield (Yobs). This is represented by the atering unit.
following equation (Semblante et al., 2014): MULESL basin was fed with the sewage coming from the preliminary
treatment stage (i.e., with the influent of the primary sedimentation),
1 1 kd SRT
¼ þ (1) whose composition is reported in Table 1, whereas MULESL outlet was
Yobs Ymax Ymax
connected to the tertiary treatment unit of the conventional WWTP.
which shows that Yobs is inversely proportional to SRT. In fact, moving Therefore, two water lines were operated in parallel at Putignano’s
from conventional activated sludge process to an extended aeration one, WWTP, the first based on the traditional primary and secondary treat­
featured by a higher SRT, allows to reduce sludge production by about ment (characterized by a plant volume of 0.26 m3/PE; 0.35 m3/PE if
30% (Sid et al., 2017). Looking at equation (1), by increasing SRT it anaerobic digestion is also included) whereas the second one on the
could be possible to reach a situation in which the biomass net pro­ innovative process having a specific plant volume of 0.19 m3/PE.
duction is close to zero (Amanatidou et al., 2016). In the conventional The main feature of MULESL system lies in the complete separation
activated sludge systems, however, this strategy could be significantly of the biomass from the liquid phase obtained in the same basin. While
hampered by the fact that the concentration of biomass cannot be biomass is confined to a dedicated zone of the basin (known as the
increased beyond a certain value due to its low settling velocity in the biomass compartment; 270 m3 volume), by means of porous plastic
secondary clarifier that would require very large sedimentation tanks. material (wheel shaped elements) packed between two metallic
Moreover, long SRT could have a negative effect on the sludge charac­ containment planes provided with suitable diffusers, the wastewater is
teristics and aeration costs for biomass suspension in the reactor (Sem­ circulated between the liquid phase compartment (400 m3), where air is
blante et al., 2014). The problem linked to the large dimensions of continuously supplied by a blower connected with membrane disc dif­
secondary settler can be solved by using a membrane biological reactor. fusers, and the biomass compartment, where the biological degradation
Nevertheless, the problems due to the high concentrations of suspended processes occur. By this way, the treated sewage is always free of
biomass, which would require frequent membrane cleaning and biomass/solids and can be easily disposed of. Biomass compartment
replacement, and high aeration flows persist (Wei et al., 2003). includes a plenum chamber for the distribution of wastewater and air,
In the present paper a new treatment system, whose acronym is above which the porous plastic material is supported (see Fig. 1).
MULESL (MUch LEss SLudge; patent n. 102017000130809), is proposed The two compartments of MULESL basin are separated by means of a
and tested at full scale for reducing the quantity of sludge in the water concrete vertical partition lower than the perimeter walls of the basin so
line of the WWTP. The potentiality of MULESL system is due to the that the liquid can overflow from the biomass to liquid phase zone (see
particular type of biomass (a mixture of self-immobilized biofilm and Fig. 1). Additionally, the two compartments are connected to each other
granular biomass), generated adopting particular operating conditions, by means of a pipe and pumping means (pump PR in Fig. 1) which allow
and retained in a porous material filling part of the reactor volume (Di the liquid to be recirculated (at flow rate of 400 m3/h) from the bottom
Iaconi et al., 2010). Granular biomass is a particular type of biofilm that of the liquid phase zone to the plenum chamber of the biomass
displays different physical and chemical characteristics compared to compartment (see Fig. 1).
activated sludge (Di Iaconi et al., 2007; Winkler et al., 2018). Thanks to In addition to the aeration provided in the liquid phase
its particular and unique physical characteristics (high density and own
shape and volume), it could be used most of the voids of the porous Table 1
plastic material thus reaching long SRTs, without the need of the sedi­ Composition (in terms of mean � standard deviation and values range) of
mentation stage, thus maximizing maintenance metabolism. Further­ municipal sewage fed to MULESL unit during the whole experimental campaign.
more, MULESL is able to simplify the water line scheme since primary Parameter Mean value (�st. dev.) Value range
and secondary treatment steps are replaced by a single stage. MULESL
pH – 6.9 (�0.4) 5.3–7.6
system can be retrofitted to existing plants by converting the activated Conductivity μS/cm 1141 (�175) 823 1590
sludge stage as well as implementing new designs. COD mg/L 1162 (�461) 480 2352
In the present paper, the performances of the first MULESL full-scale CODsol mg/L 492 (�132) 170–877
installation, obtained by retrofitting an existing activated sludge basin, BOD5 mg/L 483 (�124) 270–710
TSS mg/L 538 (�319) 162 1723
were evaluated in terms of conventional pollutants and sludge produc­
VSS mg/L 495 (�296) 100 1548
tion; the latter was compared with the one of conventional activated TKN mg/L 81.6 (�24.5) 39.4–164.0
sludge process treating the same municipal sewage. NH3 mgN/L 46.5 (�11.6) 28.0–96.2
TN mg/L 81.9 (�24.4) 39.4–164.0
Ptot mgP/L 12.3 (�4.5) 5.5–22.1
Porto mgP/L 5.9 (�2.4) 2.0–14.0
E.coli MPN/100 mL 1.1⋅108 (�0.5⋅108) 3.9⋅107 1.6⋅108

2
C. Di Iaconi et al.
Fig. 1. MULESL scheme. PR: recirculation pump; PF: feeding pump; PS: sludge extraction pump.
compartment, pure gaseous oxygen was dosed in the recirculation pipe
at a flow rate of 15 Nm3/h, in order to operate at dissolved oxygen
concentrations higher than saturation ones, thus accelerating the bio­
logical process and then improving its effectiveness. It should be noted
that pure gaseous oxygen injection takes place in a closed volume, such
as the recirculation pipe connected to the plenum chamber, thus
ensuring its complete dissolution. This allows to reduce the power de­
mand for the liquid phase compartment aeration that is known to be less
effective (conventional aeration systems usually dissolve no more than
25% of the available oxygen). MULESL basin carries out consecutive
sequences of 4 steps: feeding, recirculation, discharge and idle. During
the feeding step, sewage coming from equalization basin is sent by
means of the filling pump (at flow rate of 120 m3/h; pump PF in Fig. 1) in
the plenum chamber of the biomass compartment where it raises up to
the upper portion of the porous material, while is subjected to a filtration
with removal of suspended solids, and falls then in the liquid phase
compartment. When a predetermined level in the liquid phase
compartment is reached, also the recirculation pump is activated which
begins to recirculate the liquid from the liquid phase zone to the biomass
one, thus beginning the recirculation step, while the feeding pump
continues to fill sewage in the plenum chamber of the biomass
compartment. Therefore, feeding and recirculation steps are overlapped
for a certain time interval. When a second prefixed level in the liquid
phase zone is reached, the feeding pump stops while recirculation pump
continues to keep the liquid flowing from the liquid to the biomass
compartment (recirculation step).
During discharge step, the treated effluent (same volume fed) is
extracted from the liquid phase compartment by the same pump used for
recirculation operation (i.e., PR in Fig. 1) acting valves installed on
recirculation/discharge pipe (see Fig. 1). Finally, during idle step, the
plant is prepared to begin a new sequence of steps as the one described.
Moreover, during idle step the cleaning operation of the biomass
compartment is carried out if needed. The alternation of the treatment
sequence steps is conveniently controlled by an automation system
based on a microprocessor and a timer which manages the functioning of
the various devices interlocked to the plant: feeding pump, recirculation
pump, aeration/oxygenation system, extraction pump of the treated
effluent.
While MULESL unit continues to operate there is a continuous in­
crease in biomass concentration, due to its growth and retained sus­
pended solids present in the sewage, which at high values can lead to a
partial clogging of the biomass compartment. Increased biomass con­
centration also increases pressure of the liquid in the plenum chamber
during filling/recirculation operations, as a result of the reduction in
porous material porosity. In order to avoid such drawback, a cleaning
operation is carried out whenever the pressure in the plenum chamber
3
Journal of Environmental Management 269 (2020) 110714
during the recirculation step, measured by pressure probes, reaches 1
bar. The operation consists in flushing, during idle step, compressed air
for several minutes through a dedicated pipe provided in the plenum
chamber. The compressed air jet determines the detachment of a portion
of sludge from the plastic supporting material. The detached biomass
crosses the lower containment plane’s diffusers and settles on the bot­
tom of the plenum chamber where is extracted as liquid sludge by
activating a dedicated pump (pump PS in Fig. 1) provided with a flow
meter and sampling port in order to calculate le quantity of sludge
extracted. The extraction of the sludge continues until a certain value of
turbidity, measured at the exit of pump PS, will be reached.
2.2. Experimental campaign
MULESL unit was operated for 12 months. The experimental
campaign was split in two periods. The first period (period A), which
lasted about 7 months, referred to the transformation of activated sludge
used as inoculum in the typical biomass of the system, made up of
biofilm and granules bounded in a plastic porous. Operating conditions
of MULESL basin during this period play a decisive role for the formation
and the maintenance in time of such biomass. Experiments carried out at
laboratory scale have shown that periodicity with which wastewater is
pumped into the system, organic load applied to the biomass compart­
ment and superficial up flow liquid velocity through the porous medium
play a crucial role for obtaining and maintaining such biomass (Di Iaconi
et al., 2005, 2010). Treatment sequences of 8 h were used while the OLR
was stepwise increased up to 0.5 kg COD per m3 of biomass compart­
ment per day.
Period B was aimed at improving treatment capability once the
typical biomass was obtained. In particular, hydraulic loading to
MULESL basin was progressively increased up to 375 m3/d.
2.3. Analytical methods
MULESL plant performances were evaluated in terms of traditional
gross parameters removal efficiency and sludge production. The
following typical gross parameters were measured 2 to 3 times a week in
influent and effluent samples of MULESL unit: chemical oxygen demand
(COD), soluble chemical oxygen demand (CODsol), biochemical oxygen
demand (BOD5), total suspended solids (TSS), volatile suspended solids
(VSS), total Kjeldahl nitrogen (TKN), ammonia (NH3), nitrate (NO3 ),
nitrite (NO2 ) and total nitrogen (TN), E. coli. The latter was used for
evaluating the effectiveness of MULESL unit as disinfection system.
COD, BOD5 and TN were determined by means of Dr Lange test kits
(COD: LCK314; BOD5: LCK555; TN: LCK238). CODsol was measured
after sample filtration through a 0.45 μm filter. NH3 was measured by
C. Di Iaconi et al.
Hemera analyser whereas TSS and VSS were determined using standard
methods (APHA 2005). TKN was calculated as the difference between
TN and oxidized nitrogen. NO3 and NO2 were determined by ion
chromatography (Thermo Scientific Dionex Aquion). E. coli was detec­
ted and quantified by Colilert-18 assay according to the manufacturer’s
instructions (IDEXX, Laboratories, Inc., 2013). Colilert-18 is based on
the most probable number (MPN) procedure; thanks to the simultaneous
analysis of a large number of wells (49 large wells and 48 small wells)
and high sample volume (100 mL), it is able to provide greater accuracy
and repeatability (ISO 9308-2:2012).
The specific sludge production (SSP) of MULESL treatment was
calculated from sludge and removed COD mass balances carried out over
the 12 months of operation. In particular, it was calculated by means of
the following equation:
� � � �
� � TSSeff kgTSS
y
þTSS washing
kgTSS
y
þTSSbiomass compart
kgTSS
SSP ¼ � � shape of wheel with 9 mm height, 12 mm diameter, 800 m2/m3 specific
kgCODrem kgCOD
CODrem y
where TSSeff is the amount of suspended solids discharged with the
effluent during the year (y) of operation, TSSwashing is the amount of
suspended solids removed by washing operations during the year of
operation, TSSbiomass compart is the amount of sludge accumulated in the
biomass compartment during the experimental period , and CODrem is
the amount of COD removed during the 12 months of MULESL
operation.
TSSbiomass compart was calculated by performing, at the end of the
experimental campaign, an intensive washing operation in order to
remove completely the biomass from the porous material filling the
biomass compartment. The presence of any residual biomass in the
biomass compartment was carefully checked after the washing opera­
tion by opening the upper containment plane of the biomass compart­
ment. The detached biomass was measured in terms of volume and dry
and volatile sludge concentration. TSSeff was calculated based on hy­
draulic loading applied to the unit and effluent suspended solid content.
TSSwashing was calculated by summing the amounts of sludge removed
during the washing operations. CODrem was instead estimated from the
OLR values applied to MULESL basin during the whole period, then
multiplying each by the respective duration in days and by the relative
average COD removal efficiency.
SRT (d) of MULESL unit was calculated by the following equation:
� �
TSSbiomass compart kgTSS � �
y d
SRT ðdÞ¼ � � � � � � ⋅365
kgTSS kgTSS kgTSS y formances were evaluated in terms of conventional pollutants removal
TSSeff þTSSwashing y þTSSbiomass compart y
y efficiency and sludge production.
3. Results and discussion
3.1. MULESL plant start-up (period A)
MULESL plant was operated for 12 months. During the first 7 months
(period A) selected operating conditions were applied for generating the
typical biomass of MULESL system, consisting in biofilm and granules
bounded in a plastic porous material, starting from activated sludge
inoculum. Experiments carried out at laboratory scale revealed that the
generation of such biomass occurs in three consecutive distinct phases
(see Fig. S1). In the first phase, the activated sludge used as inoculum is
trapped in the porous material filling the biomass compartment of the
system. In the successive phase, the fraction of the activated sludge
adhered on the surface of the porous material leads to the generation of a
thin biofilm covering the surface of the filling material. The activated
sludge trapped in the interstitial pores of the filling material (both the
Journal of Environmental Management 269 (2020) 110714
inner ones of the material and the ones generated by its packing in the
biomass compartment) instead continues to grow as separated entity
(inclusion biomass). While the quantity of biomass increases, by the
increase in the biofilm thickness and in the dimensions of the inclusion
biomass, the porosity of the porous filling material is reduced, with
consequent increase of liquid rising velocity in the biomass compart­
ment. The increase of the effective rising speed due to the reduction of
the porous material provides plug-flow conditions through the biomass
compartment which have a smooth action on the inclusion biomass
particles which take the typical rounded and bevelled shape of a river
pebbles (from here the term granular biomass). Such rounded particles
reach such dimensions that they cannot go out from the pores where
they are developed (third phase), i.e. they become granules trapped in
the filling material. As expected, the features of the filling material (in
� � particular the sizes of inner and interstitial voids) have a decisive role in
kgTSS
y
holding these granules (Di Iaconi et al., 2009). Plastic elements in the
area, 0.96 g/cm3 density, 0.85 bed porosity and 120–250 mm3 voids size
were used for this aim. Yet, the rising velocity of the liquid through the
(2) biomass compartment is another important operating parameter of
MULESL systems as it affects the plug-flow conditions and hydrody­
namic shear forces, which play an important compacting action of the
biomass (Di Iaconi et al., 2005). Geometric rising speed values (defined
as the ratio between the recirculation flowrate between the liquid flow
and biomass compartments and the geometric section of the biomass
compartment) ranging between 3 and 4 m/h were used. Finally, the
feeding periodicity and OLR represent additional crucial operational
parameters since they lead to the alternation of feast and famine con­
ditions, which improve the level of biomass compactness that is vital for
the biomass concentration increase in the biomass compartment.
Treatment sequences of 8 h were used while the OLR was stepwise
increased up to 0.5 kg COD per m3 of biomass compartment per day,
obtained by a gradual increase of the volumetric loading rate. The latter
was increased, however, coherently with the effluent quality in order to
safeguard the discharge quality requirements. Therefore, during period
A, the removal efficiencies exceeded 95% for COD and TSS, and 90% for
TKN and TN, with residual concentrations in the effluent always much
lower than the discharge limits.
3.2. MULESL plant performances
Once the typical biomass of MULESL system was achieved, the plant
continued to operate for additional 5 months in order to maximize the
treatment capacity (period B). In particular, the hydraulic loading to the
plant was progressively increased up to 375 m3/d. MULESL plant per­
(3)
3.2.1. Sludge production
As reported in section 2.3, SSP was calculated by means of equation
(2). It should be noted that washing operations were never carried out
since the head loss set-point of 1 bar was never achieved while plant was
running (i.e., 12 months). Therefore, SSP was calculated taking into
account only the amount of sludge present in the biomass compartment
at the end of experimental period (i.e., TSSbiomass compart) and the sludge
lost with the effluent (i.e., TSSeff) during the 12 months of operation.
TSSbiomass compart and TSSeff turned out to be equal to 7020 and 427 kg of
dry sludge, respectively. Based on these values and applying equation
(3), a SRT of 340 d is obtained. Finally, taking into account the amount
of COD removed during the experimental period (i.e., 59,724 kg) and
applying equation (2), a SSP value of 0.13 kg TSS/kg COD removed was
obtained, corresponding to 16 gTSS/PE⋅d. This value is 3–5 times lower
than that reported in the literature for biological treatment units, based
on activated sludge process (Metcalf and Eddy Inc., 2003; Foladori et al.,
2010; Velho et al., 2019).
4
C. Di Iaconi et al. Journal of Environmental Management 269 (2020) 110714

Furthermore, the sludge production of MULESL unit was 77% lower

than that of the traditional water line of Putignano WWTP (i.e., primary
and secondary sludge) recorded during the same period (i.e., 69 gTSS/
PE⋅d). This result is even more remarkable bearing in mind that it is
obtained with a smaller volume of the water line (27% smaller; i.e., from
0.26 m3to 0.19 m3/PE).
The reduced sludge production recorded in our study is consistent
with the high SRT value of MULESL system. In fact, Vera et al. (2013)
evaluated the performances of several WWTPs operating at different
SRTs; they found that the plants based on extended aeration configu­
ration, operating at SRTs between 20 and 60 d, showed mean sludge
production values 24% lower than conventional activated sludge ones
operating at SRTs in the range 4–15 d. Furthermore, the sludge present
in the biomass compartment of MULESL unit was characterized by a
VSS/TSS ratio as low as 0.6 thus indicating that the anaerobic stabili­
zation stage may no longer be required in some cases (e.g., in the case of
landfill disposal). This would save investment costs for digester reactor
Fig. 2. COD concentration profiles of influent and effluent with COD removal
and facilities for energy production out of biogas. Compared to
efficiency of MULESL unit over the time during period B.
Putignano’s WWTP, it allows to save about 0.1 m3/PE of anaerobic
digester facility. Contrary to what obtained in our study, Amanatidou
high (higher than 94%; see Fig. 2).
et al. (2016) instead found a higher sludge VSS/TSS ratio (i.e., 0.8) in
This can be ascribed to the great stability and flexibility of the sys­
slaughterhouse WWTPs based on activated sludge process operating at
tem, whose peculiar type of biomass (a mixture of granular and biofilm
complete sludge retention. However, the stabilization degree of MULESL
confined in a porous plastic material) is able to adapt to different
sludge is coherent with the very high SRT and consistent with the
operating conditions. Yet, Fig. 3 highlights the complete removal of
non-volatile solids mass balance of the whole operating period.
BOD5 influent content throughout period B with residual effluent con­
The sludge reduction recorded in our study is significantly higher
centrations practically null.
than that reported in the literature for full scale applications of cell lysis-
MULESL plant guaranteed an excellent suspended solids removal
cryptic growth based technologies. In fact, Gardoni et al. (2011) ob­
efficiency as well, as shown in Fig. 4. In fact, a mean removal efficiency
tained 17% sludge production reduction by dosing 0.8 to 2.5 gO3/kg of
of 98% (and always higher than 93%) of the influent content was
dry solids into a fraction of 20% of the recycle sludge. Sievers et al.
recorded, even against the sharp variation in the influent value, between
(2004) have obtained sludge reduction values of up to 35% operating
162 and 1723 mg/L (see Fig. 4). It is interesting to point out the high
with ozone doses up to 6 gO3/kg of dry solids. Lower sludge reduction
stability of these performances; in fact, an effluent with a low content of
values (i.e., about 10%) were obtained by Romero et al. (2015) by using
suspended solids was produced during the whole experimental period
ozone doses between 0.7 and 5 gO3/kg of volatile solids. Velho et al.
with mean concentration values of 8 mg/L and always lower than 16
(Velho et al., 2016, 2019) operating at full scale with an anaerobic
mg/L (see Fig. 4).
side-stream reactor have reported sludge production reduction values
Influent VSS/TSS ratio was always higher than 0.7 (very often higher
between 20% and 28% compared with conventional activated sludge
than 0.85), highlighting that suspended material in the influent was
configuration. Furthermore, it should be considered that these tech­
almost organic, also in accordance with the COD and CODsol (approxi­
nologies lead to an increase of the plant reaction volume, and then of
mately 50% of COD content of the influent was ascribable to particulate
investment costs, since an additional reactor is inserted in the return
material in the sewage).
activated sludge line of the biological compartment. Velho et al. (2016)
The remarkable removal efficiencies of the MULESL plant in terms of
obtained at Levico WWTP a 20% reduction in sludge production by
suspended solids should be attributed to its biomass compartment: it
inserting an anaerobic side stream reactor in the return activated sludge
operatively becomes, in effect, a very low porosity filter in which the
line of the biological unit. This led to a 32% increase in reaction volume
biomass works as a filtering medium. Once trapped in the biomass,
of the biological compartment. The reduced sludge production was
suspended material is hydrolyzed by the enzymes therein and
increased up to 30% by increasing the volume of anaerobic side stream
reactor. However, this approach increased the biological unit volume by
47%.

3.2.2. Plant performances in terms of the traditional gross parameters

MULESL performances during period B were recorded in terms of
influent and effluent time-concentration profiles, and relative removal
efficiencies of COD, BOD5, TSS, TKN, NH3 and TN.
Looking at Fig. 2, it is possible to observe that MULESL guaranteed a
mean COD removal of 97% (always higher than 87%) with an average
residual concentration in the effluent of 33 mg/L (always lower than 73
mg/L), hence well below the most restricted discharge limit (i.e., 100
mg/L) set by the Italian regulation and inforce for Putignano WWTP. It is
worth noting that these performances were obtained regardless of the
volumetric organic load applied to the plant, which was even as high as
3.3 kgCOD per m3 of biomass compartment per day.
Yet, it should also be pointed out that these high removal efficiencies
were obtained in the face of great variability of influent COD concen­
tration. COD value entering MULESL basin has greatly fluctuated even
reaching a maximum value of 2400 mg/L (see Fig. 2); COD removal Fig. 3. BOD5 concentration profiles of influent and effluent with BOD5 removal
efficiency in this borderline case was however found to be considerably efficiency of MULESL unit over the time during period B.

5
C. Di Iaconi et al. Journal of Environmental Management 269 (2020) 110714

Fig. 4. TSS concentration profiles of influent and effluent with TSS removal Fig. 6. NH3 concentration profiles of influent and effluent with NH3 removal
efficiency of MULESL unit over the time during period B. efficiency of MULESL unit over the time during period B.

subsequently metabolized. This peculiarity provides the plant highly

rugged performances.
Concerning nitrogen, Figs. 5 and 6 show that MULESL unit was able
to ensure a high efficiency in removing TKN and NH3 (the latter ac­
counts for about 50% of the first one).
This highlights the existence of a nitrification process, which appears
to be very stable. In fact, mean removal efficiencies of 95% (and always
higher than 88%) for TKN and NH3 were recorded, with mean effluent
concentrations of only 3.5 and 1.9 mg/L (and always lower than 7.3 and
5.9 mg/L), respectively. Yet, these performances were obtained in the
face of great variability of the influent values of the parameters (i.e.,
from 39.4 to 164.0 mg/L for TKN and from 28.0 to 66.0 mg/L for NH3;
see Figs. 5 and 6).
Furthermore, MULESL unit was also able to ensure an excellent total
nitrogen removal efficiency by means of the complete denitrification
process, although the treatment sequence carried out by the plant did
not include a final planned anoxic step (i.e., a phase during which ox­
ygen is not supplied). In fact, TN concentration values out from the plant Fig. 7. TN concentration profiles of influent and effluent with TN removal ef­
were always extremely low and identical to those of TKN (which, as ficiency of MULESL unit over the time during period B.
known, represents the sum of ammonia and organic nitrogen), thus
pointing out the complete reduction of any oxidized nitrogen species. (Metcalf and Eddy Inc., 2003).
The profiles reported in Fig. 7 show mean TN removal efficiencies of The profiles of the nitrogenous species (i.e., TKN, oxidized forms and
95% (always higher than 88%) with mean effluent concentration of 3.5 ammonia) within a typical treatment sequence have shown that nitri­
mg/L (always lower than 7.4 mg/L). These performances are even more fication and denitrification processes take place at the same time: in
relevant since they were obtained at very low excess sludge production. fact, NH3 (both that already present in the sewage and that formed by
In fact, it is well know that in the traditional activated sludge process a ammonification process of organic nitrogen) is simultaneously oxidized
substantial amount of nitrogen is removed with the waste sludge stream to nitrite and/or nitrate and then reduced to molecular nitrogen, which,
being a gas, abandons the reaction environment.
The simultaneous nitrification-denitrification process can be
ascribed both to the high concentration of biomass present in the
biomass compartment and to the dynamic operating conditions (typical
of sequential reactors), which generate, inside the biomass (especially in
the granular one), external aerobic and internal anoxic layers, where
nitrification and denitrification take place, respectively.
MULESL effectiveness in removing pathogens from wastewater was
evaluated on the basis of E.coli, which represents a bacterial pathogen
surrogate (Thwaites et al., 2018). It is the most used microbiological
indicator of water quality and to simulate the pathogen’s behavior in
WWTPs (Seeger et al., 2016). MULESL unit was able to remove 2.5 to 2.7
log units of E.coli, with residual effluent concentrations of 4.4⋅104 to
7.2⋅104. These performances are in line with those of primary and sec­
ondary treatments of WWTPs that usually remove about 2–3 log units of
E. coli with residual concentrations in secondary effluents in the range of
104–105 CFU/100 mL (Russo et al., 2019). However, it should be borne
in mind that MULESL system assured such performances in a single step.
Fig. 5. TKN concentration profiles of influent and effluent with TKN removal
Finally, according to the low sludge production value obtained,
efficiency of MULESL unit over the time during period B.

6
C. Di Iaconi et al.
MULESL unit showed low phosphorus removal efficiencies as well (i.e.,
on average, 12%). Removal efficiencies as high as 40% were also
recorded during some days, which can be ascribed to the filtering
capability of MULESL towards particulate phosphorus.
4. Conclusions
A new treatment system, whose acronym is MULESL (MUch LEss
SLudge), was proposed and tested at full scale for reducing the quantity
of sludge in the water line of the sewage treatment plant. MULESL unit
for 3500 PE, fed with the influent of the primary sedimentation of
Putignano’s WWTP, was monitored for 1-year in terms of sludge pro­
duction and typical gross parameters removal. Solid and COD mass
balances carried out over the whole operating period gave a specific
sludge production value of 0.13 kg of dry sludge per kg of COD removed.
This value was 77% lower than that obtained for primary and secondary
treatments of Putignano’s WWTP during the same period. Furthermore,
MULESL sludge was already stabilized, being characterized by an
organic matter content of 60%. MULESL unit guaranteed high and stable
performances with mean removal efficiency higher than 95% for COD,
BOD5, TSS, TKN, NH3 and TN Finally, MULESL unit also acted as a good
disinfection system ensuring 2.5 to 2.7 log units removal of E.coli with
effluent concentrations of 4.4⋅104 to 7.2⋅104.
Declaration of competing interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
CRediT authorship contribution statement
Claudio Di Iaconi: Conceptualization, Supervision, Data curation,
Writing - original draft. Marco De Sanctis: Investigation, Writing -
original draft. Valerio Guido Altieri: Data curation, Investigation.
Acknowledgment
This work was partially supported by the Italian National PON
TARANTO Project (ARS01_00637) and MANGIAFANGHI project
(“Sludge reducing during municipal wastewater") funded by Puglia re­
gion (Italy) within “Regional Technological clusters” call -FSC 2007/
2013.
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://doi.
org/10.1016/j.jenvman.2020.110714.
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