Freezing/thawing effects on anaerobic digestion of mixed sewage sludge

A. Montusiewicz
*
, M. Lebiocka, A. Ro
_
zej, E. Zacharska, L. Pawowski
Institute of Environmental Protection Engineering, Faculty of Environmental Engineering, Lublin University of Technology, ul. Nadbystrzycka 40B, 20-618 Lublin, Poland
a r t i c l e i n f o
Article history:
Received 30 September 2009
Received in revised form 11 December 2009
Accepted 19 December 2009
Available online 27 January 2010
Keywords:
Freezing/thawing pre-treatment
Sludge disintegration
Sludge solubilization
Mixed sewage sludge
Enhanced biogas yield
a b s t r a c t
This study examined the effects of freezing/thawing disintegration on the characteristics of mixed (pri-
mary and waste) sewage sludge from municipal wastewater treatment plants. It also assessed the effects
of freezing/thawing on anaerobic digestion, and its consequences for biogas production and digester
supernatant quality. Freezing/thawing caused a decrease of more than 10% in the total chemical oxygen
demand (COD), total nitrogen (TN), volatile solids (VS) and total solids (TS). A simultaneous doubling of
the soluble COD and volatile fatty acids (VFA) occurred. Release of nitrogen and phosphorus compounds
was also found.
The biogas yield obtained from frozen/thawed sludge was 1.31 m
3
kg
1
of removed VS; this exceeded
approximately 1.5 times the value for the raw sludge.
On the basis of the global mass balance it was indicated that freezing/thawing of the mixed sewage
sludge followed by anaerobic digestion could be considered as a double-phase digestion rather than
a pre-treatment method.
2010 Elsevier Ltd. All rights reserved.
1. Introduction
Current thinking is that anaerobic digestion of sewage sludge
should focus both on highly efcient sludge stabilization and on
enhanced biogas production. Operational data relating to munici-
pal wastewater treatment plants (WWTPs) have shown that anaer-
obic digesters work with insufcient loads and possible capacity
reserves reach as much as 30% of the digesters volume (Braun,
2002). This has particularly been shown at high ow rates. In such
cases, co-fermentation of sewage sludge and other organic compo-
nents could be introduced to improve biogas generation (Mont-
usiewicz, 2008). Another fairly recent technological advancement
that enhances anaerobic digestion efciency and increases biogas
productivity has been the development of pre-treatment tech-
niques that accelerate the hydrolysis of sludge (Elliott and Mah-
mood, 2007). Sludge pre-treatment incorporates different
disintegration methods, such as mechanical (e.g., sonication,
hydrodynamic cavitation), chemical (e.g., alkali treatment, ozona-
tion), thermal (e.g., heat treatment, freezing/thawing) and biologi-
cal (e.g., treatment by enzymes) (Erdinclerm and Vesilind, 2000;
Saktaywin et al., 2005; Xue and Huang, 2007; Jan et al., 2008).
Some of these methods are thought to improve the dewatering
capability. However, most pre-treatments for municipal activated
sludge lead to cell disruption (Bien et al., 2004) that promotes sol-
ubilization of organic matter and releases intracellular and cell
wall polymers (including polysaccharides, proteins, lipids and
other macromolecules) into the liquid phase. Interestingly, differ-
ent mechanisms are responsible for membrane damage. The
hydrolysis of sludge can be accomplished by its exposure to wet
milling with small beads (mechanical disintegration), or to high-
temperature and high-pressure gradients resulting from the rapid
collapse and expansion of microbubbles (sonication, hydrody-
namic cavitation), or to highly oxidative conditions provided by
ozone, which reacts with components of cell membranes and con-
verts them into smaller molecular-weight compounds (Elliott and
Mahmood, 2007). A high-temperature treatment sterilizes the
sludge, which splits the cells due to pressure differences during
the hydrolysis process (Keep et al., 2000). Treatment by enzymes
incorporates primary and secondary cell lysate (formed in lysate-
thickening centrifuges), which contains some enzymes, parts of en-
zymes and cofactors that accelerate cellular degradation and sup-
port bacterial growth (Dohnyos et al., 1997). Freezing causes the
formation of intracellular crystals, which mechanically damage cell
membranes (Thomashow, 1998).
Freezing/thawing processes frequently occur in nature, leading
to changes in soil characteristics (Eigenbrod, 1996). The composi-
tion of sewage sludge that is treated and dumped outside in cold
regions could also vary as a result of freezing (Rush and Stickney,
1979; Reed et al., 1986). In municipal wastewater treatment plants
freezing/thawing could be used for highly efcient dewatering
(Martel, 1989). Sewage sludge dewatering by freezing is accom-
plished through the separation of solid and liquid fractions during
the formation of ice crystals. It is stated that freezing promotes
transformation of the ocs into a more compact form (Chu et al.,
1999; Jean et al., 2001). However, to obtain the best results, the
0960-8524/$ - see front matter 2010 Elsevier Ltd. All rights reserved.
doi:10.1016/j.biortech.2009.12.125
* Corresponding author. Tel.: +48 81 538 4325; fax: +48 81 538 1997.
E-mail address: A.Montusiewicz@wis.pol.lublin.pl (A. Montusiewicz).
Bioresource Technology 101 (2010) 34663473
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sludge must be completely frozen at a relatively slow rate since
benecial effects of colder temperatures, longer periods of freezing
and slower freezing rates were noticed (Vesilind and Martel, 1990;
rmeci and Vesilind, 2001; Wang et al., 2001). According to Reed
et al. (1986), sludge freezing may convert a non-drainable jelly-like
consistency into a granular material that drains immediately when
it thaws. This is particularly important for sludge that contains
alum due to its extremely low dewatering capability by gravity
drainage (Martel (2000), the aggregated particles found in frozen/
thawed alum sludge are described as coffee ground). Precondi-
tioning allows the achievement of more than 20% of dry mass in
sludge as soon as thawing is completed. Moreover, some further
drying provides concentration of solids of up to 50%, which is not
possible using mechanical devices.
Most studies concerning the mechanisms of freezing/thawing
processes and their inuence on the qualitative characteristics of
the product relate to waste-activated sludge and alum sludge (Ves-
ilind et al., 1991; rmeci and Vesilind, 2001). When sludge is fro-
zen, both the suspended and dissolved solids are rejected by a
growing ice front (Vesilind et al., 1991; Martel, 2000). This dis-
placement gathers the solids into larger particles. As a result of
freezing, the sludge is being converted to a matrix of ice crystals
and aggregated solid particles. Martel (2000) found differences in
morphology of the ice crystals. He reported that the alum sludge
ice crystals grow in columns. Whereas growth of the activated
sludge ice crystals becomes dendritic (dendrites denote branching
tree-like structures), which was attributed to the presence of dis-
solved solids in activated sludge. Water surrounding the particles
trapped in the ice creates a very thin layer, commonly called a
transition layer, that does not freeze at normal temperatures.
Vesilind et al. (1991) stated that the transition layer is more easily
replenished when the particles are small.
Chu et al. (1997), similar to Kawasaki and Matsuda (1995),
investigated the effect of electrolyte (sodium chloride) addition
on the subsequent freezing/thawing of the excess activated sludge,
and thus on its dewater ability, settleability and residual moisture.
The authors concluded that addition of sodium chloride retards the
sludge particles migration and remarkably improves the sludge
lterability.
rmeci and Vesilinds studies (2001) regarding activated sludge
and alum sludge indicated that freezethaw conditioning causes
cell disruption and release of intracellular material to the sludge
supernatant. In activated sludge the process mentioned strongly
increases the concentration of proteins, carbohydrates and cations
in the supernatant, however in alum sludge a noticeable release is
not observed. The authors concluded that to improve activated
sludge dewater ability the concentration of proteins, carbohy-
drates and cations should previously be decreased.
The literature review by Prez-Elvira et al. (2006) shows that
the inuence of sludge pre-treatment technology preceding its
anaerobic digestion on the enhanced biogas production, sludge re-
moval efciency and pathogen reduction was so far investigated
regarding different methods, although the freezing/thawing tech-
nique was examined only with reference to the dewatering ability
(Reed et al., 1986; Martel, 1989; Chu et al., 1997; rmeci and Ves-
ilind, 2001; Wang et al., 2001). Its involvement in anaerobic diges-
tion seems to be counter-productive due to high energy
consumption. This does not apply to some countries that have spe-
cic climate conditions, or that have access to natural or articial
ice thereby allowing freezing without additional operational
costs. The practical aspects of this process and its effect on the
anaerobic stabilization of sewage sludge should be examined. If
the production of biogas could be increased, the whole process
would become more cost-effective despite environmental
restrictions.
Only a few studies regarding sludge pre-treatment via freezing/
thawing in the enhancement of its anaerobic digesting efciency
were found; furthermore, they concern merely waste-activated
sludge (Wang et al., 1995). Reports relating to the evaluation of
the effects of such pre condition technique on the chemical compo-
sition of mixed sewage sludge (which consists of primary sludge
and waste-activated sludge) were not found. Similarly, its inu-
ence on anaerobic digestion process and biogas production was
not investigated. Considering a specic character of the primary
and waste-activated sludge and taking into account that mixed
sludge is a typical digester feed in the full-scale systems, these is-
sues are worth researching.
In the present study, the inuence of freezing/thawing disinte-
gration on the change in characteristics of mixed sewage sludge
was evaluated. Moreover, its effect on anaerobic digestion was
analyzed based on changes in bioreactor operational conditions,
enhanced biogas production and the quality of the digester super-
natant. The freezing/thawing technique followed by the mixed
sludge anaerobic digestion was evaluated on the basis of the global
mass balance calculations and the question of whether freezing/
thawing disintegration could be considered as a pre-treatment
method or as the rst stage of the double-phase digestion.
2. Methods
2.1. Material characteristics
Sewage sludge that included two-source residues was obtained
from the Puawy municipal WWTP, Poland, using primary and sec-
ondary treatments. Sludge originating from a gravity thickener (i.e.
primary thickened sludge) and from a mechanical belt thickener
(i.e. waste thickened sludge) was used as material for our study.
The characteristics of thickened sludge from Puawy WWTP are
shown in Table 1. It should be noted that wastewater from the
meat industry, which supplied the municipal sewer system, inu-
enced the sludge characteristics, especially with regard to the total
COD value (high levels were observed).
2.2. Sample preparation procedure
Sludge was sampled daily in the Puawy municipal WWTP and
then provided immediately to the laboratory of the Lublin Univer-
sity of Technology (Poland). Sludge from a gravity thickener and
from a mechanical belt thickener was transported in separate con-
tainers. Under laboratory conditions, sludge was mixed at a vol-
ume ratio of 60:40 (primary:waste sludge), then homogenized
and partitioned. Such samples were considered to be raw sludge
(RS). Some of the samples were disintegrated using a freezing/
thawing technique. The sludge was frozen at 25 C for 24 h in a
laboratory freezer and then thawed for another 12 h at 20 C in
the indoor air. Freezing/thawing conditions were selected similarly
to Jan et al.s studies (2008); however, some modications were
applied. The preliminary experiments showed that the mixed
sludge froze completely at 25 C and at least 12 h were required
to completely thaw the samples at room temperature. The as-
sumed operational conditions complied with these results. The
samples prepared using the freezing/thawing procedure were con-
sidered to be frozen/thawed sludge (FTS).
2.3. Laboratory installation of sludge anaerobic digestion
The laboratory installation consisted of two anaerobic digestion
systems operating in parallel. Each of them included a completely
mixed, hermetic reactor with a working volume of 40 dm
3
inserted
into the heating jacket at a stable temperature and equipped with
A. Montusiewicz et al. / Bioresource Technology 101 (2010) 34663473 3467
the required instrumentation. Mixing was carried out using a
mechanical stirrer that has a rotational speed of 50 min
1
. Bioreac-
tor equipment included an inuent peristaltic pump feeding the di-
gester, as well as storage vessels both for inuent and efuent
(digested medium). At the top of the reactor, an attachment for
biogas installation was tted. The gas system consisted of pipelines
linked with the pressure equalization unit and the drum gas meter.
This was equipped with gas valves, a dewatering valve and a gas
sampler with a rubber septum, which enabled insertion of a syr-
inge with a pressure lock. The laboratory installation is shown in
Photo 1.
2.4. Operational set-up
An inoculum for the laboratory reactors was taken from the
Puawy WWTP as a collected digest from a mesophilic anaerobic
digester operating at 3537 C with a volume of 2500 m
3
and a
hydraulic retention time of about 18 days. The adaptation of the di-
gester biomass in the systems under investigation was achieved
after 30 days.
The experiment was carried out simultaneously in two parallel
systems operating at a controlled mesophilic temperature of 35 C.
The semi-ow reactors were supplied regularly once a day with 2 L
of feed sludge. The reactor RS (control) was fed using raw sludge,
whereas the reactor FTS (experimental) was supplied using fro-
zen/thawed sludge. The hydraulic retention time reached 20 days
and the hydraulic loading rate was 0.05 d
1
for both reactors. How-
ever, the organic loading rate (OLR) differed for the RS and FTS
reactors and was 1.521.62 kg VS m
3
d
1
for the RS system and
1.271.35 kg VS m
3
d
1
for the FTS system, respectively. The study
was run over 60 days.
2.5. Analytical methods
For raw and frozen/thawed sludge, the following parameters
were analyzed each day: total chemical oxygen demand (COD),
volatile fatty acids (VFA), total solids (TS), volatile solids (VS), total
nitrogen (TN), alkalinity, sludge density and pH level. The same
schedule was used for determining the values of parameters that
characterized supernatant (sludge liquid phase) before digestion:
soluble COD (SCOD), total nitrogen in supernatant (TNs), ammonia
nitrogen (NNH

4
), nitrite and nitrate nitrogen (NNO

x
) and ortho-
phosphates (PPO
3
4
). The supernatant samples were obtained by
centrifuging the sludge at 4000 r min
1
for 30 min.
In digested sludge, specied parameters were determined three
times a week, in accordance with the assumed timetable. Similarly,
the supernatant of digested sludge was examined using the same
schedule.
Most analyses were carried out in accordance with the proce-
dures in the Polish Standard Methods. Some analyses were per-
formed with FIASTAR 5000 using FOSS analytical methods:
ammonium was determined according to ISO 11732, nitrite and ni-
trate according to ISO 13395 and ortho-phosphate in accordance
with ISO/FDIS 156811.
The anaerobic digestion efciency was controlled by the daily
evaluation of biogas yield and its composition (CH
4
, CO
2
and other
gases). Biogas production was determined using Drum-type Gas-
meter TG-Series (Ritter, Germany). The composition of the biogas
was measured using a gas chromatograph Shimadzu GC 14B cou-
pled with a thermal conductivity detector (TCD) tted with
glass-packed columns. The Porapak Q column was involved to
determine CH
4
and CO
2
concentrations. The parameters used for
the analysis were as follows: injector 40 C, column oven 40 C,
detector 60 C and current bridge 150 mA. The carrier gas was he-
lium with a ux rate of 40 cm
3
min
1
. Peak areas were determined
using the computer integration program (CHROMA X).
3. Results and discussion
3.1. Effect of freezing/thawing on sludge characteristics
The characteristics of raw and frozen/thawed sludge that sup-
plied, respectively, the RS (control) and FTS (experimental) system
are listed in Table 2 (average data are reported).
In comparison with the raw sludge, a 12% decrease in the aver-
age total COD concentration was observed for the frozen/thawed
medium. Simultaneously, a decline in TS, VS (Table 2) and TN (Ta-
ble 3) values of 16.1%, 16.9% and 15.1%, respectively, were shown.
There was a more than twofold increase in SCOD compared with
the average concentration as a result of freezing/thawing disinte-
gration. Concomitantly, a rapid rise in VFA concentration (more
than double), as well as a simultaneous drop in pH value to 5.82,
was observed. An increase in alkalinity (as CaCO
3
) from 1100 to
1400 mg L
1
and a decrease in sludge density from 1008 kg m
3
to 1006 kg m
3
were also noticed. The above results indicated a
relationship between the increase in soluble contaminants and,
at the same time, the decrease in total organic matter in frozen/
thawed sludge. A high concentration of both SCOD and VFA and
Table 1
Characteristics of thickened raw sludge from the Puawy (Poland) municipal wastewater treatment plant during 2008.
Value Primary thickened sludge Waste thickened sludge
TS
g kg
1
VS
% (of TS)
TS
g kg
1
VS
% (of TS)
Average 30.35 75.31 60.85 76.47
Upper/Lower 95% mean 32.45/28.24 77.11/73.51 66.81/54.88 78.03/74.92
Photo 1. Laboratory installation of sludge anaerobic digestion.
3468 A. Montusiewicz et al. / Bioresource Technology 101 (2010) 34663473
a simultaneous decrease in pH level seems to conrm the solubili-
zation of organic matter and the release of intracellular material.
This observation is consistent with the research by Stabnikova
et al. (2008) regarding food waste and rmeci and Vesilind
(2001) regarding activated sludge. Moreover, another study simi-
larly reported that freezing/thawing disintegration caused an in-
crease in the soluble fraction of carbohydrates in increased SCOD
during the treatment of waste sludge from food-processing waste-
water (Jan et al. 2008). The same authors indicated that a critical
value of pH (5.5) was required for initiation of the methanogenic
stage, however according to Vavilin et al. (2001) only hydrogeno-
trophic methanogenesis is suspected to be possible at such acidic
pH. In the presented research, the average value of pH was 5.82,
thus the boundary requirements for the initiation of the methano-
genic stage were fullled.
In the present study the complex mechanisms of freezing/thaw-
ing disintegration could lead to the formation of the specic buffer-
ing conditions. However, the increase of the VFA concentration was
higher than alkalinity changes which, as a consequence, caused the
pH decrease.
It is known that the biological effects of biomass freezing are
dominated by destruction of living cells (resulting from direct
mechanical action of ice crystals) or by damaging them via changes
in the composition of the liquid phase (e.g. concentration of the
solutes) (Pegg, 2007). On the other hand, it is commonly accepted
that freezing causes lethal effects for some microorganisms. How-
ever, recent studies relating to the freezing of food have indicated
that some microorganisms (even those that are considered to be
sensitive for inactivation) are only inhibited at low temperatures
(Kennedy, 2000; Miladi et al., 2008). It is worth noticing that some
food components, such as proteins or fats, tend to protect the
microorganisms and act as cryoprotectants (e.g. glycerol). More-
over, it is stated that a combination of slow freezing and rapid
thawing could favour the survival of the microorganisms (Ken-
nedy, 2000).
The research by rmeci and Vesilind (2001) regarding activated
sludge indicated that freezethaw conditioning caused cell disrup-
tion and release of intracellular material to the sludge supernatant.
Additionally, they reported the start of anaerobic reactions in the
activated sludge for 12 h of thawing. Following this, the authors
propose a similar explanation for the mixed sludge and suggest
that the COD, TS, VS and TN losses, and the observed solubilization,
could result from two possible mechanisms that might take place
simultaneously, even though one of them could prevail. The rst
mechanism involves the reaction of both the immobilizing exoen-
zymes present in the system and the endoenzymes released from
the cells disrupted as an effect of extracellular and intracellular
ice formation. The second approach suggests that biomass activity
could not be excluded and degradation could be carried out by the
microorganisms that survived in frozen sludge. The presence of the
cryoprotectant components (e.g. proteins and fats) in the raw sew-
age sludge could have an inuence on the microorganisms, pro-
tecting them from freezing, thus biodegradation lasting for 12 h
during the thawing stage could occur.
The analysis of the effects of freezing on nutrient concentration
seems to conrm sludge solubilization. The concentration of nutri-
ents in raw and frozen/thawed sludge that supplied, respectively,
the RS and FTS systems is listed in Table 3. An average reduction
in TN affected by sludge freezing was found at approximately
143 mg L
1
. However, an average concentration of TNs in the
sludge supernatant showed almost a twofold increase for the fro-
zen/thawed sludge, thus indicating the nitrogen release from the
biomass. Moreover, higher concentrations of ammonia nitrogen
and ortho-phosphates were also observed. Their values increased
by approximately 37 mg L
1
for ammonia nitrogen and by 99 mg
L
1
for phosphorus as ortho-phosphates. Such observations are
Table 2
Characteristics of raw (RS) and frozen/thawed sewage sludge (FTS) supplied to RS and FTS systems.
Parameter Unit Value Raw sludge (inuent to RS reactor) Frozen/thawed sludge (inuent to FTS reactor)
pH Average 6.39 5.82
Upper/Lower 95% mean 6.46/6.32 5.87/5.77
Alkalinity mg L
1
Average 1100 1400
Upper/Lower 95% mean 1157/1043 1470/1330
COD mg L
1
Average 43,844 38,566
Upper/Lower 95% mean 45,531/42157 42,586/34,546
SCOD mg L
1
Average 3190 6756
Upper/Lower 95% mean 3411/2969 7638/5874
VFA mg L
1
Average 1757 3755
Upper/Lower 95% mean 1804/1710 3845/3665
TS g kg
1
Average 41.0 34.4
Upper/Lower 95% mean 42.3/39.7 35.5/33.3
VS g kg
1
Average 31.4 26.1
Upper/Lower 95% mean 32.3/30.5 26.9/25.3
Table 3
Concentration of nutrients in raw and frozen/thawed sewage sludge.
Parameter Unit Value Raw sludge (RS reactor) Frozen/thawed sludge (FTS reactor)
TN mg L
1
Average 950 806.7
Upper/Lower 95% mean 1119.7/780.3 1031.4/582.0
TNs (supernatant) mg L
1
Average 190 340
Upper/Lower 95% mean 288.7/91.3 419.2/260.8
NNH

4
(supernatant) mg L
1
Average 94.0 130.9
Upper/Lower 95% mean 99.7/88.3 154.9/106.9
PPO
3
4
(supernatant)
mg L
1
Average 86.4 185.2
Upper/Lower 95% mean 100.4/72.4 199.6/170.8
A. Montusiewicz et al. / Bioresource Technology 101 (2010) 34663473 3469
consistent with results obtained by Xue and Huang (2007) during
thermal treatment of excess sludge. The authors found that heating
the sludge at a temperature of no less than 50 C led to the efcient
release of not only phosphorus and nitrogen compounds, but also
organics and some metal cations.
Considering the similarity of the freezing and thermal treat-
ment effects (which were, in part, conrmed in our investigation),
it could be expected that some metal cations would be released
and would increase the alkalinity of the sample. Such an increase
was observed during our experiment in frozen/thawed sludge that
buffered acidication resulting from an increase in VFA concentra-
tion and a decrease in pH level. It should be noted that the concen-
trations of nitrite and nitrate nitrogen were negligible during all
experiments that were undertaken.
3.2. Effect of freezing/thawing on operational conditions and biogas
yields
Table 4 includes essential values of loading rates in the systems
under investigation. It should be noted that the hydraulic loading
rate and hydraulic retention time were the same in both reactors
and were 0.05 d
1
and 20 d, respectively.
Based on the differences in the characteristics of raw and disin-
tegrated sludge, presented and discussed before, the higher loading
rates relating to VS and COD were estimated in the RS reactor (con-
trol) and compared to the FTS reactor. However, loading rates eval-
uated for the FTS reactor with regard to SCOD and VFA exceeded
more than twice the values for the RS digester, which was in accor-
dance with a higher concentration of solutes via freezing. The VFA/
VS inuent ratios were 0.056 and 0.144, respectively, in the sys-
tems supplied with raw (RS) and frozen/thawed (FTS) sludge.
It is commonly accepted that a higher organic loading rate
(according to Tchobanoglous et al. (2003), a value of 4.8 kg m
3
d
1
should not be exceeded) and a higher concentration of VS fa-
vours more efcient anaerobic digestion, whereas a decrease of
such parameters diminishes biodegradation effects. This was con-
rmed during the present studies on the basis of removal of VS
at the level of 51.9%, with the average OLR being 1.57 kg VS m
3
d
1
, and of 44.5% with an OLR value of 1.31 kg VS m
3
d
1
, respec-
tively, in the RS and FTS reactors.
In the investigated systems, however, various loading rates did
not inuence the parameters of digested sludge. Results showed
that efuent parameters, such as TS and VS (Table 5), and TN (Ta-
ble 7, presented and discussed later in section 3.3), were similar in
both treatments and the value differences amounted only to 6.8%
for TS, 4.0% for VS and 4.6% for TN. A greater difference of 45.8%
was found for SCOD as a twofold lower concentration was ob-
served in the FTS reactor probably due to microorganism adapta-
tion for solute uptake. Moreover, high alkalinity (above 4000 mg L

1
) and low VFA concentration (not exceeding 300 mg L
1
) and a
favourable pH value of 7.37.4 indicated that a methanogenic
phase was stable. Comparable parameters were achieved despite
the differences in feed sludge that supplied reactors and the vari-
ous loading rates (see Table 4).
The average biogas yields attained during investigations are
shown in Fig. 1. The biogas production per kg of VS removed, which
was calculated in the RS system and amounted to 0.86 m
3
kg
1
VS,
was in compliance with the range of 0.751.12 m
3
kg
1
VS that was
suggested by Tchobanoglous et al. (2003) as an adequate yield
from sewage sludge anaerobic digestion. A comparison of such val-
ues in both systems showed that the biogas yield obtained from
the frozen/thawed sludge exceeded approximately 1.5 times the
value achieved from the raw sludge and reached 1.31 m
3
kg
1
of
volatile solids removed. A similar tendency was observed with re-
gard to the biogas yields based on the removed TS and COD. The
increased values were achieved in the FTS system despite a lower
solids loading rate and a minor concentration of VS. The possible
explanation is that the solubilization of organic matter resulted
from sludge disintegration and this was conrmed by both the
high concentrations of SCOD and VFA. An increased content of
the solutes regarded as the substrate, being more available and
easy to convert for microorganisms, resulted in an improved biogas
yield.
During experiments, a comparable biogas composition in both
reactors was observed (Fig. 2). The average methane content was
61.5% and 63.0%, respectively, in the system fed with raw and fro-
Table 4
Operating conditions for anaerobic digestion.
Reactor Value Loading rate(kg m
3
d
1
) refers to:
VS COD SCOD VFA
RS Average 1.57 2.19 0.16 0.088
Upper/Lower 95% mean 1.62/1.52 2.28/2.10 0.17/0.15 0.09/0.086
FTS Average 1.31 1.93 0.34 0.19
Upper/Lower 95% mean 1.35/1.27 2.13/1.73 0.38/0.30 0.195/0.185
Table 5
Efuent quality of the anaerobic digestion process.
Parameter Unit Value Reactor RS Reactor FTS
pH Average 7.73 7.84
Upper/Lower 95% mean 7.87/7.59 8.06/7.62
Alkalinity mg L
1
Average 4275 4350
Upper/Lower 95% mean 4304/4246 4411/4289
COD mg L
1
Average 23,734 19,130
Upper/Lower 95% mean 25,256/22,212 20,574/17,686
SCOD mg L
1
Average 2342 1270
Upper/Lower 95% mean 2407/2277 1337/1203
VFA mg L
1
Average 263 217
Upper/Lower 95% mean 379/147 288/146
TS g kg
1
Average 24.9 23.2
Upper/Lower 95% mean 25.7/24.1 23.8/22.6
VS g kg
1
Average 15.1 14.5
Upper/Lower 95% mean 15.3/14.9 15.2/13.8
3470 A. Montusiewicz et al. / Bioresource Technology 101 (2010) 34663473
zen/thawed sludge; an average difference of only 1.5% was ob-
tained. These results indicated that disintegration of mixed (pri-
mary and waste) sewage sludge via freezing/thawing inuenced
the share of biogas components to a minor extent.
The average methane yields achieved during the studies are
presented in Fig. 3. It should be noticed that higher methane pro-
duction was found from frozen/thawed sludge in comparison to
raw sludge. The observed increases were 36.1%, 14% and 37.6%,
respectively, with regard to the removed VS, COD and TS. Wang
et al. (1995) found, similarly, that methane generation was im-
proved from waste-activated sludge that was pre-treated by freez-
ing; however, a lower increase of about 27% was achieved with
regard to VS removed. Providing feedstock for anaerobic digestion
consisting of mixed sludge instead of waste-activated sludge could
explain a higher increase in methane yield in the present study.
Methane is a source of energy for producing electricity and heat,
and it is especially protable in terms of enhancing its generation
from sewage sludge.
To verify results achieved during experiments, the calculations
of mass balance in the form of measurable parameters were under-
taken for the organic material uxes (that entered and left the
reactors). According to Grady et al. (1999), COD should be used
as a parameter for organic material measurement in an anaerobic
process related to methane generation. Arajo et al. (1998) pro-
posed an extended approach and calculated the COD/VS ratio as
a constant that reected the conversion of VS into COD mass.
Moreover, to convert methane into COD mass, it was considered
that there is a stoichiometric relationship of 4 mg COD mg
1
CH
4
(Grady et al., 1999) and that 1 L of CH
4
at 35 C represents a COD
mass of 64/25.3 = 2.53 g.
The mass balance in the digester, based on a COD mass in such
uxes as an inuent (MS
inf
), an efuent (MS
eff
) and methane
(MS
CH
4
), was expressed according to Arajo et al. (1998) as follows:
- COD in digester inuent
MS
inf
Q COD
inf
f
cv
VS
inf
1
- COD in digester efuent
MS
eff
Q COD
eff
f
cv
VS
eff
2
- COD in methane
MS
CH
4
2:53 Q
CH
4
3
It should be noted that COD
inf
and COD
eff
values refer to SCOD con-
centration and the f
cv
value expressed the COD/VS ratio for both
examined treatments.
Arajo et al. (1998) proposed the f
cv
constant value of 1.5 mg
COD mg
1
VS for waste-activated sludge. However, in our study,
based on a mixed sludge (primary sludge was also included), the
values of COD/VS ratio were determined using the experimental
data both for supplied and digested medium. The data required
and results achieved from mass balance are shown in Table 6.
The percentage fraction of recovered organic material in the di-
gester (included in Table 6) was calculated according to the
equation:
B MS
eff
MS
CH
4
=MS
inf
100 4
The calculations of mass balance were based on experimental
data and were therefore subject to errors. Unimportant differences
were found between the uxes of organic material entering the
reactors and leaving them. These differences amounted to 2.3%
and 3.2% for the RS (control) and FTS digester, respectively. This
shows them to reect normal measurement uncertainty and leaves
the 14% increase of methane yield per COD removed in FTS a
remarkable value for all the mass balance errors.
The results of mass balance calculations exclude the effects of
sludge transformations during freezing/thawing pre-treatment.
However, the sludge solubilization and acidication, as well as
the concurrent losses of 12% for COD and 17% for VS noticed after
freezing/thawing process, indicate conversion of the organic com-
pounds into soluble forms or their degradation. Therefore, the
freezing/thawing process followed by anaerobic digestion could
be considered as a double-phase digestion rather than a pre-
treatment method, thus the whole process should be involved in
the global digestion calculations. The results obtained using such
an approach showed similar values for the biogas yields per kg of
VS removed, respectively of 0.9 m
3
kg
1
and 0.86 m
3
kg
1
for the
frozen/thawed sludge and the raw one. The rst value was calcu-
lated concerning the double-phase digestion and quantifying
the volatile solids that were previously converted to SCOD and
VFA in frozen/thawed sludge; the second one was obtained for
conventional system (sludge anaerobic digestion without pre-
treatment RS). The biogas yields calculated per VS fed were also
Fig. 1. The biogas yields obtained from RS (raw sludge) and FTS (frozen/thawed)
systems.
Fig. 2. The biogas composition observed in RS and FTS systems.
Fig. 3. The methane yield achieved from RS and FTS systems.
A. Montusiewicz et al. / Bioresource Technology 101 (2010) 34663473 3471
comparable in both systems; the achieved values reached 0.49 m
3
kg
1
in the double-phase process and 0.45 m
3
kg
1
in the con-
ventional digestion. It is worth noticing that the removal of VS
reached 53.8% for the combined process and was slightly higher
than the level of 51.9% observed in the conventional one. Global
mass balance carried out for the removed COD gave the biogas
yield of 0.62 m
3
kg
1
for the double-phase digestion and 0.7
for the conventional system. To sum up, from a global point of view
the effects of the combined process were similar to the conven-
tional one.
3.3. Supernatant quality
The digester supernatant characteristics are presented in Ta-
ble 7. Comparing data from Table 3 and Table 7, a higher release
efciency was achieved in the RS reactor due to the nutrients re-
leased directly from raw sludge (without disintegration). The ob-
served concentrations of released TNs, NNH

4
and PPO
3
4
in the
supernatant increased approximately 2.3 times for TNs, almost 8
times for NNH

4
and 1.6 times for PPO
3
4
in comparison to the
supernatant composition of the raw sludge. Lower efciency was
attained in the FTS reactor, as some nutrients were released previ-
ously during the freezing/thawing pre-condition. In this case, the
concentration of released TNs and NNH

4
in the digester superna-
tant increased almost 1.4 times and 6 times, respectively, in com-
parison to the supernatant of the FTS reactor. However, a small
decrease in PPO
3
4
concentration (to about 52 mg L
1
) was ob-
served in the reactor that was fed with frozen/thawed sludge.
It was found that, despite the differences in nitrogen and phos-
phorus released from freezing/thawing of sewage sludge (shown in
Table 3), the characteristics of the digester supernatant were com-
parable in both systems. Moreover, the observed NNH

4
values
were similar to those seen by Song et al. (2004) during mesophilic
anaerobic digestion of sewage sludge. The authors found that the
concentration was 630 mg L
1
, whereas during our investigations
observed levels in supernatants were, respectively, 750.1 mg L
1
from the RS reactor and 775.6 mg L
1
from the FTS reactor. For
PPO
3
4
concentrations, much lower values were achieved by Song
et al. (42.4 mg L
1
) compared to our observations at the levels of
137.2 mg L
1
and 133.6 mg L
1
, respectively, in the RS and FTS
supernatant.
The comparable NNH

4
concentrations obtained during our re-
search could result from either of the following: total nitrogen loss
during freezing (Table 3) and a higher struvite (MgNH
4
PO
4
) precip-
itation in the FTS reactor. The latter explanation is in accordance
with a lower PPO
3
4
concentration (Table 7) and a higher pH value
(Table 5) in the FTS supernatant.
It can be concluded that freezing/thawing of the mixed sewage
sludge did not inuence the digester supernatant quality. Such
observations are essential as the supernatant quality is an impor-
tant factor that could inuence the nutrient mass balance during
advanced wastewater treatment (Battistoni et al., 1998). The high
loads of ammonia nitrogen and ortho-phosphates that supplied
the wastewater treatment system in the supernatant stream could
affect its overloading and a subsequent decrease of nutrient re-
moval efciency and an increase of total treatment cost.
4. Conclusions
The freezing/thawing disintegration altered the mixed sewage
sludge characteristics. Decreases in the COD, TN, TS and VS values
and simultaneously more than twofold increases in both the SCOD
and VFA concentrations were observed. A release of nitrogen and
phosphorus compounds also occurred.
The average biogas yield from frozen/thawed sludge reached
1.31 m
3
kg
1
of removed VS and exceeded approximately 1.5 times
the value from the raw sludge.
On the basis of the global mass balance it was indicated that the
freezing/thawing of the mixed sewage sludge followed by anaero-
Table 7
Concentration of nutrients in digested sludge.
Parameter Unit Value RS efuent FTS efuent
TN mg L
1
Average 756.7 793.3
Upper/Lower 95% mean 821.0/692.4 912.2/674.4
TNs (supernatant) mg L
1
Average 426.7 466.7
Upper/Lower 95% mean 491.0/362.4 540.3/393.1
NNH

4
(supernatant) mg L
1
Average 750.1 775.6
Upper/Lower 95% mean 786.6/713.6 840.4/710.8
PPO
3
4
(supernatant)
mg L
1
Average 137.2 133.6
Upper/Lower 95% mean 148.8/125.6 144.4/122.8
Table 6
COD mass balance for anaerobic digesters data and results.
Parameter Unit Reactor
RS FTS
Inuent Efuent Inuent Efuent
Mass balance data
Q L d
1
2 2 2 2
SCOD g L
1
3.19 2.34 6.76 1.27
VS g kg
1
31.4 15.1 26.1 14.5
Sludge density kg m
3
1008 1002 1006 1002
COD/VS 1.4 1.6 1.5 1.3
Q
biogas
L d
1
28.35 30.75
CH
4
in biogas % 61.5 63.0
Q
CH4
L d
1
17.44 19.37
Mass balance results
MS
inf
(Eq. (1)) g COD d
1
95.00 92.29
MS
eff
(Eq. (2)) g COD d
1
53.10 40.32
MS
CH4
(Eq. (3)) g COD d
1
44.11 49.01
COD mass balance (Eq. (4)) % 102.3 96.8
3472 A. Montusiewicz et al. / Bioresource Technology 101 (2010) 34663473
bic digestion could be considered as a double-phase digestion
rather than a pre-treatment method.
Acknowledgements
Authors thank the nancial support from the Ministry of Sci-
ence and Higher Education of Poland, PBZ-MEiN-3/2/2006.
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