ARTICLE IN PRESS

International Biodeterioration & Biodegradation 57 (2006) 88–92

www.elsevier.com/locate/ibiod

Biological activity during composting of sewage

sludge at low temperatures
Rosa Margesin, Jan Cimadom, Franz Schinner
Institute of Microbiology, Leopold Franzens University, Technikerstrasse 25, A-6020 Innsbruck, Austria
Received 30 October 2005; accepted 1 December 2005
Available online 19 January 2006

Abstract

The feasibility of aerobic composting of sewage sludge under cold climate conditions in the European Alpine winter season was studied
using a 40:60 (v/v) mixture of sewage sludge from a municipal wastewater treatment plant and shrub clippings. Despite ambient air
temperatures that ranged from a maximum of 10.1 to
0.2 1C to a minimum of 2.2 to
3.6 1C, the temperature increased to 66 1C in the
core of the pile. One compost pile was turned at a higher frequency (twice a week) than a second pile. This resulted not only in a loss of
heat, but also in statistically significant greater desiccation (po0.001) of the composting material than in the second pile. This in turn had
a significantly negative impact on microbial processes, as shown by poor decrease in the C/N ratio, reduced enzyme activities, respiration
and biomass, and a high level of stress response (qCO2) and toxicity (inhibition of fresh mass production of Lepidium sativum). The
significant negative correlation between the dry mass content of the compost material and several biological parameters demonstrated
the sensitivity of microbial activity towards desiccation. The study indicates that low moisture content is a more limiting factor for
composting than low temperature.
r 2005 Elsevier Ltd. All rights reserved.

Keywords: Sewage sludge; Composting; Moisture; Enzymes; Respiration; Cold climate

1. Introduction To date, most of the research on composting has focused

Sewage sludge, a product of waste water treatment, is
rich in nutrients and trace elements and could be re-used in
agriculture as fertilizer and soil conditioner. High odour
emission, high levels of heavy metals and toxic organic
compounds, and the presence of potentially pathogenic
microorganisms, demand pretreatment of sewage sludge
before application in agriculture (Veeken and Hamelers,
1999; Tiquia et al., 2002a). Composting is a successful
strategy for the sustainable recycling of organic wastes
(Fermor, 1993; Tuomela et al., 2000). Surprisingly, there is
little information on aerobic composting of sewage sludge,
and the available information is mainly dedicated to the
characterization of humic and fulvic acids resulting from
organic matter transformation (Amir et al., 2005a,b,c;
Jouraiphy et al., 2005).
Corresponding author. Tel.: +43 512 5076021; fax: +43 512 5072929.
E-mail address: rosa.margesin@uibk.ac.at (R. Margesin).
on changes in physico-chemical parameters, primarily in an
effort to find a reliable indicator of compost maturity
(Mathur et al., 1993; Amir et al., 2005b). Comparatively
little information is available on biological and biochemical
parameters as an indicator for process rate evaluation,
although the usefulness of biological parameters as
indicators of microbial activity and dynamics during
composting has been demonstrated (Lei and Vander-
Gheynst, 2000; Vuorinen, 2000; Tiquia et al. 2002a,b;
Mondini et al., 2004). There is, however, very limited
knowledge about the role of enzymes and other biological
parameters during the composting of sewage sludge.
Composting has been shown to be feasible under
temperate climate conditions, at ambient air temperatures
around 20–30 1C (Tiquia et al., 2002a,b). Considerably
lower temperatures prevail in winter in temperate climates
or throughout the year in cold climates. Composting
under low temperature conditions is thought not to be
practicable because of low microbial degradation activities.
McCartney and Eftoda (2005) observed the impact of

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doi:10.1016/j.ibiod.2005.12.001
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R. Margesin et al. / International Biodeterioration & Biodegradation 57 (2006) 88–92
snowfall on oxygen supply and pile moisture content, and
the impact of large frozen zones on pathogen inactivation
efficiency. Others reported successful composting of muni-
cipal sludge (Smith, 1984) or animal manure (Lynch and
Cherry, 1996) at ambient temperatures of
15 to
28 1C.
The objective of this study was to investigate the
feasibility of aerobic sewage-sludge composting in an
Alpine region in winter at mean monthly air temperatures
around 0 1C, using piles that were subjected to different
turning frequencies. Biological activities as well as the most
important physico-chemical parameters were monitored.
2. Materials and methods
2.1. Composting
Two piles were built in a hangar in winter time (January). The piles had
a trapezoid shape, 40 m in length, 4 m in width at the base and 2 m in
height, and had a volume of approx. 200 m3. The starting material was the
same in both piles and consisted of 40% (v/v) sewage-sludge from a
municipal wastewater treatment plant and 60% (v/v) shrub clippings in
order to add a carbon source and to favour aerobic microbial
transformation. Aeration was ensured by forced ventilation during the
first 3 weeks of composting, and by an extraction fan during the residual
incubation time.
One pile (A) was covered with a 5-cm layer of mature compost to
insulate it and minimize water loss. The cover also acted as a biofilter to
minimize odour emission. This pile was first turned 3 weeks after set-up
(whereby the layer was mixed into the composting material), and
afterwards once each week using a compost turner. The second pile (B)
was turned twice each week from the beginning, and therefore could not
be covered with mature compost.
Five 2-kg samples (sub-samples were mixed homogeneously to give
composite samples) were collected from the core of each pile at day 1 and
then weekly (on days 6, 14, 20, 28, 34, and 48). The five sampling positions
were located 80 cm from the base of the piles and at a distance of 12, 16,
20, 24 and 28 m from one end. Immediately after collection, samples were
transported in cool boxes to the laboratory, stored at 4 1C, and analyzed
within 4 days. The temperature in the core of the piles was recorded by
temperature sensors inserted at the sampling positions.
2.2. Chemical and physical analyses
Field-moist samples, sieved to o10 mm, were used to determine dry
mass (105 1C, 24 h) and pH. The pH was determined with a glass electrode
in a mixture of 10 g composting material and 50 ml distilled water. The
C/N ratio was calculated from contents of total organic carbon (TOC) and
total nitrogen (Kjeldahl) in air-dried samples (20 1C, 2 days) ground to
o1 mm. The methods were as described by Schinner et al. (1996) and
prescribed in Austria (BGBl II, 2001).
2.3. Biological analyses
Biological analyses were carried out on field-moist samples, and results
expressed on a dry mass basis. Enzyme activities, viz. xylanase, protease,
urease and fluorescein diacetete (FDA) hydrolytic activity, basal respira-
tion (CO2-evolution), substrate-induced respiration (as an indirect
measure of microbial biomass), and the metabolic quotient qCO2 (the
ratio CO2:C produced by basal respiration/biomass C, were determined as
described by Schinner et al. (1996) and Margesin (2005) using optimized
masses of compost material. Toxicity was evaluated by determining the
above-ground fresh mass of Lepidium sativum (cress) using 15% of
composting material in the test assay (Österreichisches Normungsinstitut,
1993).
89
2.4. Statistical data treatment
All analyses were carried with two to three replicates per sample, and
the mean results per sample used for statistical data treatment. Statistical
calculations were carried out using the software Statistica 6.1. Normal
distribution of the data was tested by the Shapiro–Wilks’ W test.
Significant differences between the two treatments were analyzed by the t-
test for independent groups (data with normal distribution) or by the
Mann–Whitney U-test (non-parametric data). Treatments were considered
significant when po0.05. Correlations between the measured parameters
were analyzed by Spearman rank order correlation (non-parametric data)
or Pearson product-moment correlation (normal distribution).
3. Results and discussion
3.1. Chemical and physical parameters
At the start of the composting process the mean
temperature in the core of the two piles was 1473 1C.
Maximum and minimum ambient air temperatures during
the running of the piles ranged from 10.1 to
0.2 1C and
2.2 to
3.6 1C, respectively (Fig. 1). Temperature increased
in both piles with time and was significantly higher
ðpo0:001Þ in pile A than in pile B over the whole
incubation period (Table 1). Clearly, the high turning
frequency resulted in a loss of heat. Maximum temperature
values were measured after 34 days, and were 6674 1C in
A and 4875 1C in B, indicating the development of
thermophilic microbial communities. After 48 days, the
temperature had decreased to 5674 1C in A and 4471 1C
in B. The incubation period was too short to attain a
mesophilic temperature level. Unfortunately, it was not
possible to run the piles for longer. For economic reasons
set by our industrial partner, the composting time generally
takes 7 weeks, after this period the composting material is
sieved and subjected to post-maturation outdoors.
A temperature of 55–60 1C is required for pathogen
reduction, but this criterion was met in pile A. Other
studies on low-temperature composting have reported pile
temperatures of 55 1C after 15–20 days (Smith, 1984), 50 1C
70
60
50
Temperature (°C)
40
Pile A
30
Pile B
20 Air (max.)
Air (min.)
10
0
-10
0 10 20 30 40 50
Days
Fig. 1. Changes in ambient air temperatures and pile (core) temperatures
during composting.
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Table 1
Effect of frequent turning on physical, chemical and biological parameters during composting of sewage sludge in piles A (turned at a low frequency) and
B (frequently turned) over an incubation period of 48 days and at the end of the treatment

Parameter All sampling periods ðn ¼ 30Þ After 48 days of treatment ðn ¼ 5Þ

Mean or median value (%) Significantly different? Mean or median value (%) Significantly different?

Pile A Pile B Pile A Pile B

Temperature 393 287 Yes 408 321 Yes

Dry mass 81 127 Yes 81 132 Yes
pH 101 98 No 94 91 Yes
C/N 82 124 Yes 83 103 Yes
Respiration 63 33 Yes 49 14 Yes
Biomass 46 14 Yes 37 7 Yes
FDA hydrolytic activity 106 80 Yes 109 78 Yes
qCO2 119 196 Yes 125 262 Yes
Xylanase 103 85 Yes 147 90 Yes
Protease 474 219 Yes 203 111 Yes
Urease 100 96 No 23 39 No

Mean or median values (depending on the presence or absence of normal distribution of the data) are relative percentage values (t0 ¼ 100%).
 po0.001.
 po0.01.
 po0.05.

after 12–26 days (Lynch and Cherry, 1996) or above 65 1C
at ambient temperatures below
20 1C (McCartney and
Eftoda, 2005). In comparison, a maximum temperature of
72 1C was obtained after 4 days during composting of
sewage sludge and straw (Amir et al., 2005a) and green
waste (Jouraiphy et al., 2005) in a warm climate
(Morocco). The maximum temperature during sludge
composting under warm ambient conditions was then only
higher by 6 1C than the maximum in the present study
under cold conditions, but the lag time was significantly
higher under cold conditions.
Moisture content is an important parameter influencing
biological activity and biochemical rates. The starting
material had a dry mass content of 4073%. Over the
whole incubation period, the material of pile A was
characterized by a statistically significantly lower moisture
level (po0.001) than pile B, which was turned more
frequently (Table 1). After 48 days, the dry mass content
had decreased to 3270.4% in pile A, but had increased to
5375% in pile B. Clearly, the high turning frequency (pile
B) resulted in a higher desiccation of the compost material.
This, however, had a significantly negative impact on
microbial activity (for details, see 3.2).
The pH value also affects the composting process. The
starting material had a pH of 7.170.3. This remained
almost constant over most of the incubation period in both
piles owing to the high buffer capacity of the sewage sludge
components, but decreased slightly to pH 6.7 and 6.5 in
piles A and B, respectively, after 48 days. These values were
within the optimum pH range for bacteria and fungi. In
comparison, Amir et al. (2005a) measured a pH of 6.2 in
final compost of activated sludge.
During efficient composting, the C/N ratio is expected to
decrease as a consequence of degradation of organic matter
and mineralization. Frequent turning of the composting
material (pile B) resulted in significantly reduced degrada-
tion due to low microbial activity (see 3.2), as shown by the
almost unchanged C/N ratio over time. The difference
between the two treatments was statistically significantly
different (Table 1). The C/N ratio decreased in pile A from
an initial 25.8 to 21.7 in 48 days. At that time, the
composting process, however, was not completed. Prolon-
gation of the experiment would have led to a significantly
lower C/N ratio. Other authors reported a C/N decrease
from about 30 to 12.2 after 135 days (Amir et al., 2002a), to
16.4 after 180 days (Amir et al., 2002c), and to 12 after 135
days (Jouraiphy et al., 2005) during composting of sewage
sludge with straw or fresh green plant waste. The small
decrease in the C/N ratio in our study may also be
attributed to sewage sludge components, such as low
molecular fatty acids resulting form anaerobic sludge
treatment, which inhibited microbial metabolism.
A significant negative correlation between temperature
and C/N values was noted during composting of the
sewage sludge (Table 2). This indicates that a large
temperature increase is of crucial importance for efficient
mineralization, which in turn results in reduced C/N
values.
3.2. Biological activity
In general, biological parameters tested in this study
indicated unfavourable conditions in pile B. Consequences
of the high frequency of turning, i.e. heat loss and
desiccation, had a negative impact on microbial activity.
All forms of microbial activity tested, except urease, were
significantly higher (po0.01–0.001) in the composting
material of pile A than in that of pile B (Table 1).
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Table 2
Correlation coefficients and significance levels between physical, chemical and biological parameters during composting of sewage sludge in pile A over an
incubation period of 48 days (n ¼ 30)

Dry mass pH C/N Respiration Biomass FDA hydrolytic activity Xylanase Protease Urease

Temperature
0.353 NS 0.117 NS
0.490
0.250 NS
0.306 NS 0.337 NS 0.436 0.372 NS
0.202 NS
Dry mass
0.527 0.758
0.435 0.065 NS
0.395
0.292
0.447 0.474
pH
0.663 0.700 0.371 0.303 NS
0.243 NS 0.382
0.157 NS
C/N 0.490 0.637
0.521
0.551 0.284 NS
0.020 NS
Respiration 0.789
0.181 NS
0.554 0.169 NS 0.111 NS
Biomass
0.037 NS
0.383 0.246 NS 0.175 NS
FDA hydrol. act. 0.229 NS 0.044 NS
0.060 NS
Xylanase 0.054 NS
0.286 NS
Protease
0.110 NS

NS, not significant.

po0.001.
po0.01.
 po0.05.

Respiration measured in composting material is a
measure of the total biological activity and results from
the degradation of organic matter; the formation of CO2 is
the last step of carbon mineralization. The time course
during composting depends on the starting material
(Mondini et al., 2004). In the present study, both basal-
and substrate-induced respiration (as an indirect parameter
of microbial biomass) decreased after 20 days (pile A) or
from the beginning (pile B). In pile A, these two parameters
were higher by a factor of 2 (basal respiration) and 3
(biomass) than in pile B. Consequently, the metabolic
quotient qCO2 was significantly higher in pile B. The qCO2
is an indication of the efficiency of compost microorgan-
isms and is often seen as a stress response (Sparling, 1997;
Margesin et al., 2000). The higher the value, the less
efficient are microbial processes (Schinner et al., 1996).
The applicability and usefulness of enzyme activity
measurements in monitoring and characterizing the com-
posting process has been reported (Tiquia et al., 2002c;
Mondini et al., 2004). Enzyme activities measured in the
present study generally increased with time (as composting
progressed) in pile A, indicating that the indigenous
microorganisms were able to synthesize the enzymes
required for degradation and hydrolysis of various organic
compounds, including complex substances. The potential
of litter and wood decomposition is reflected by xylanase
activity, since this enzymes is one of the most important in
primary litter degradation. In this study, xylanase activity
had significantly increased only after 34 days, by which
time easily available compounds were diminished. There
was a significant positive correlation between xylanase
activity and temperature (Table 2), showing that elevated
temperatures during the thermophilic phase of composting
are essential for rapid degradation of complex organic
compounds, such as wood components (Tuomela et al.,
2000). Protease activity had a much shorter lag time than
xylanase, owing to the early release of proteins during the
degradation of organic matter. A similar time course was
observed with FDA hydrolytic activity, which is a
summation parameter of overall hydrolytic activity. In
the composting material of pile B, however, no relative
increase of enzyme activities was detected. Urease activity
also showed a tendency to increased values in pile A
compared with pile B; a significant difference, however,
was not detected, because of high standard deviations.
The significant negative correlation between the dry
mass content and several biological parameters (Table 2)
demonstrates the sensitivity of microbial activity, especially
of enzymes, to desiccation, and indicates that low moisture
content is a factor more limiting for composting than low
temperature. It also demonstrates the importance of
measuring enzyme activities in moist samples rather than
in air-dried samples.
After composting for 48 days, the toxicity of the material
obtained was determined by measuring above-ground fresh
mass of L. sativum seedlings. An inhibitory effect
(80710%) was detected for material from pile B, whereas
material from pile A stimulated growth by 2576%. This
demonstrates that growth-inhibiting substances that were
initially present in sewage sludge were eliminated in the
course of composting in pile A. Owing to reduced
microbial activity in pile B, inhibitory compounds could
not be eliminated efficiently, but accumulated instead.
Altogether, despite the fact that for logistical reasons, we
were unable to run the composting process long enough to
achieve a stable compost material, the data show the
feasibility of composting sewage sludge under the cold
winter conditions prevailing in the European Alpine
region. However, running the composting process at low
temperature will need to take into account the moisture
content, which is a major factor influencing the process.
Since the rise in temperature is slower for composting
processes run under cold conditions, strategies such as pile
coverage or reduction of the frequency of pile turning will
need to be considered in choosing the strategies for
optimising the process.
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Acknowledgments
This study was supported by the DAKA Entsorgungs-
unternehmen GmbH & Co KG. We thank A. Haeusler and
M. Klingler for running the piles, E. Rudel (Zentralanstalt
fuer Meteorologie und Geodynamik) for providing us with
meteorological data.
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