Review

TRENDS in Biotechnology Vol.19 No.9 September 2001

363

Challenge of psychrophilic anaerobic

wastewater treatment
Gatze Lettinga, Salih Rebac and Grietje Zeeman
Psychrophilic anaerobic treatment is an attractive option for wastewaters that
are discharged at moderate to low temperature. The expanded granular sludge
bed (EGSB) reactor has been shown to be a feasible system for anaerobic
treatment of mainly soluble and pre-acidified wastewater at temperatures of
510C. An organic loading rate (OLR) of 1012 kg chemical oxygen demand
(COD) per cubic meter reactor per day can be achieved at 1012C with a
removal efficiency of 90%. Further improvement might be obtained by a
two-module system in series. Stabile methanogenesis was observed at
temperatures as low as 45C. The specific activity of the mesophilic granular
sludge was improved under psychrophilic conditions, which indicates that
there was growth and enrichment of methanogens and acetogens in the
anaerobic system. Anaerobic sewage treatment is a real challenge in moderate
climates because sewage belongs to the complex wastewater category and
contains a high fraction of particulate COD. A two-step system consisting of
either an anaerobic up-flow sludge bed (UASB) reactor combined with an
EGSB reactor or an anaerobic filter (AF) combined with an anaerobic hybrid
reactor (AH) is successful for anaerobic treatment of sewage at 13C with a
total COD removal efficiency of 50% and 70%, respectively.

Gatze Lettinga
Salih Rebac*
Grietje Zeeman
Sub-department
Environmental
Technology, Wageningen
University,
Bomenweg 2,
6703 HD Wageningen,
The Netherlands.
*e-mail: Salih.Rebac@
Algemeen.mt.wag-ur.nl

Particularly under temperate climate conditions,

many low and medium strength wastewaters are
discharged at low ambient temperatures, including
domestic wastewater and a large variety of industrial
wastewater, such as those from bottling, malting,
brewery and soft drinks manufacturing plants. Such
industrial processes might produce several streams
with different characteristics either in flow or in
chemical oxygen demand (COD) concentrations, thus
some wastewater might have a broad concentration
range because the COD of industrial effluents
depends largely on the technological process.
Low strength industrial wastewaters generally
contain a variety of biodegradable compounds such
as simple short-chain volatile fatty acids (VFAs),
alcohols and carbohydrates, but frequently also
contain proteins, suspended solids of different origin,
fats or long-chain fatty acids (LCFAs).
Moreover, many wastewaters belong to the
category very low strength, that is with COD
values in the range 0.3 to 0.6 g dm3, and sometimes
even lower, as in domestic sewage. In the case of
sewage, the pollution also consists of relatively
complex compounds and during winter periods in
moderate climates its temperature might
temporarily drop to 510C.
Effect of low temperature on anaerobic biological
conversion

Microorganisms are classified into temperature

classes on the basis of the optimum temperature
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and the temperature span in which the species are

able to grow and metabolize (Fig. 1). The
overlapping growth temperature ranges in Fig. 1
indicate that there isnt a clear boundary between
these classic groups of psychrophilic, mesophilic and
thermophilic microorganisms. The bacterial growth
rates of methanogenic thermophiles and mesophiles
from anaerobic reactors are well determined.
However, to date, only one psychrophilic
methanogenic bacteria from a permanent cold
environment (the Ace lake in Antarctica)1, two
psychrotrophic marine methanogens2 and a few
psychrotrophic acetogenic bacteria (homoacetogens)
from natural sediments have been isolated3,4. The
fact that anaerobic psychrophiles have merely been
found in natural ecosystems illustrates the lack of
information about anaerobic reactors treating
wastewater under psychrophilic conditions.
Under psychrophilic conditions, chemical and
biological reactions proceed much slower than under
mesophilic conditions. Most reactions in the
biodegradation of organic matter require more
energy to proceed at low temperatures than at a
temperature optimum of 37C (Table 1). However,
some reactions, such as hydrogenotrophic sulfate
reduction, hydrogenotrophic methane production
and acetate formation from hydrogen and
bicarbonate, require less energy (Table 1; reactions 9,
10 and 11, respectively).
A strong temperature effect on the maximum
substrate utilization rates of microorganisms has
been observed by many researchers818. In general,
lowering the operational temperature leads to a
decrease in the maximum specific growth and
substrate utilization rates but it might also lead to an
increased net biomass yield (g biomass g 1 substrate
converted) of methanogenic population12,19 or
acidogenic sludge20.
Effect of low temperature on the physical and chemical
properties of wastewater

A drop in temperature is accompanied with a change

of the physical and chemical properties of the
wastewater, which can considerably affect design and
operation of the treatment system. For instance, the
solubility of gaseous compounds increases as the
temperature decreases below 20C. This implies that
the dissolved concentrations of methane, hydrogen
sulfide and hydrogen will be higher in the effluent of
reactors operated at low temperatures than those

0167-7799/01/$ see front matter 2001 Elsevier Science Ltd. All rights reserved. PII: S0167-7799(01)01701-2

Review

364

Growth rate methanogens (%)

Fig. 1. Relative growth

rates of psychrophilic,
mesophilic and
thermophilic
methanogens after
Wiegel5.

TRENDS in Biotechnology Vol.19 No.9 September 2001

100

follows that the diffusion constant of soluble

compounds is ~50% lower at 10C compared with the
mesophilic temperature range (3040C).

Thermophiles

80

Anaerobic wastewater treatment technology

60

Mesophiles

40
Psychrophiles

20
0
0

20

40
60
Temperature (C)

80

TRENDS in Biotechnology

from reactors operated at high temperatures. The

high increase of solubility of CO2 indicates that a
slightly lower reactor pH might prevail under
psychrophilic conditions.
At low temperatures, the viscosity of liquids is
also increased. Therefore, more energy is required
for mixing and sludge bed reactors become less
easily mixed, particularly at low biogas production
rates. In psychrophilic reactors, particles will settle
more slowly because of a decreased liquidsolid
separation at low temperatures. Moreover, related
to the higher liquid viscosity, the diffusion of soluble
compounds will drop at lower temperatures, as
indicated in Eqn 1 (Ref. 21).
T
D2 = D1 1 2
T1 2

(1)

In Eqn 1, D = diffusion coefficient of a specific

compound (m2 s1), T = temperature (K), and = the
liquid viscosity of the solution (N s m2). The
subscripts 1 and 2 refer to two different temperatures.
The data in Table 2 provide diffusivity of soluble
compounds in the temperature range 10 to 40C,
relative to the diffusivity at 30C. From Table 2 it
Table 1. Stoichiometry and Gibbs free-energy changes of acetate,
propionate, butyrate and hydrogen anaerobic conversion in the presence
and absence of sulfate
G kJ
(37C)

Reactions

reaction1
(10C)

CH3CH2COO + 3H2O CH3COO + HCO3 + H+ + 3H2

+71.8

+82.4

CH3CH2COO + 0.75SO42 CH3COO + HCO3 + 0.75HS + 0.25H+ 39.4

35.4

CH3CH2COO + 1.75SO42 3HCO3 + 1.75HS + 0.25H+

88.9

80.7

CH3CH2CH2COO + 2H2O 2CH3COO + H+ + 2H2

+44.8

+52.7

CH3CH2CH2COO + 0.5SO42 2CH3COO + 0.5HS + 0.5H+

29.3

25.9

CH3CH2CH2COO

128.3

116.4

CH3COO + SO42 2HCO3 + HS

49.5

45.3

CH3COO + H2O CH4 + HCO3

32.5

29.2

4H2 + SO42 + H+ HS + 4H2O

148.2

157.1

10

4H2 + HCO3 + H+ CH4 + 3H2O

131.3

140.9

11

4H2 + 2HCO3 + H+ CH3COO +4H2O

98.7

111.8

2.5SO42

4HCO3

2.5HS

0.5H+

Energy changes were calculated by using the Van t Hoff equation, standard enthalpy values of
compounds6 and Gibbs free-energy changes7 at 25C.

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One of the major successes in the development of

anaerobic wastewater technology was the
introduction of high-rate reactors in which biomass
retention and liquid retention are uncoupled2325.
This feature comprises a crucial issue for the
treatment of low(er) strength wastewater. In some
reactor systems, the sludge retention is based on the
settling characteristics of sludge aggregates and
thickening and expansion characteristics of the
established flocculent sludge bed with its inherent
structures, such as the well-known Upflow
Anaerobic Sludge Bed (UASB) reactors (Fig. 2). In
such systems, the hydraulic load will become the
restrictive factor with respect to the required
reactor volume when treating very low strength
wastewater.
Obviously, the low CODinfluent will result in low
substrate levels (50100 mg COD l1) inside the
reactor and also in a low biogas production rate. In
conventional anaerobic sludge bed reactors, this
implies low mixing intensities in the reactor and
consequently in a poor substratebiomass contact.
Another serious problem when treating very low
strength wastewater is that the permissible amount
of sludge washout per cubic meter of wastewater is
extremely small, which sets exceptionally high
requirements on the sludge retention abilities of the
reactor. Therefore, the required reactor volume in
the case of treating low strength wastewater will
generally be determined by the permissible hydraulic
loading rate (HLR) rather than by the organic loading
rate (OLR) (Ref. 26). Practically all full-scale
applications of anaerobic wastewater treatment so
far are restricted to wastewater with temperatures
exceeding 18C.
Psychrophilic anaerobic treatment of wastewater

The established sanitary wastewater-engineering

world presently considers anaerobic wastewater
treatment of cold and very low strength wastewater,
not to be practically feasible. This opinion is mainly
on the basis of prejudice and a serious lack of insight
into the anaerobic digestion process and technology.
In fact, in the past, only a few scientists have
undertaken research in this field9,14,27,28. Most
studies with the effect of temperature on anaerobic
digestion show a strong negative effect on the
metabolic activity of mesophilic anaerobic
methanogenic bacteria at decreasing temperature,
as presented in Fig. 3. This indicates that the
capacity of an anaerobic reactor seeded with
mesophilic biomass will drop sharply during start-up.
Temperature effects on maximum substrate
utilization as shown in Fig. 3 can be described
mathematically using the Arrhenius equation30.

Review

TRENDS in Biotechnology Vol.19 No.9 September 2001

365

Table 2. The diffusivity of any soluble compounds at various temperature

relative to the diffusivity at 30Ca
Temperature (C)

10

20

30

40

50

60

Dtemp/D 30 ()

0.57

0.77

1.00

1.26

1.55

1.88

were calculated using eqn 1 and the viscosity of pure water at the various temperatures21.

Because temperature strongly affects the rates of

the anaerobic conversion processes, some essential
improvements have to be made in the conventional
design of high-rate reactors to enable their
application under sub-optimal temperatures and for
very low strength wastewater. If successful, such a
modified reactor system would represent a major
technological break-through, and efficient
bioengineering of bacterial catalysis under
sub-optimal temperatures would be possible.
Such a successful application of psychrophilic
anaerobic biocatalysis would be of great
economical importance because, in general
(depending on the temperature of the wastewater),
a significant amount of energy is required to
bring the wastewater temperature up to the
optimal mesophilic range (3040C; Ref. 31). This
puts a heavy burden on the economy of the
wastewater system.
The feasibility of high-rate anaerobic wastewater
treatment (AnWT) systems for cold wastewater
depends primarily on: (1) the quality of the seed
material used and its development under submesophilic conditions; (2) an extremely high sludge
retention under high hydraulic loading conditions
because little if any viable biomass can be allowed to
wash out from the reactor; (3) an excellent contact
between retained sludge and wastewater to utilize
all the available capacity within the bioreactor;
(4) the types of the organic pollutants in the
wastewater32; and (5) the reactor configuration,
especially its capacity to retain viable sludge.
Single and multi-compartment (moduled)
reactors can be considered for the configuration of
the reactor. To accomplish the highest possible
overall treatment efficiency (especially in the case
of multi-component wastewater) moduled reactors,
namely the staged sludge anaerobic reactors
(SSARs), offer significantly better potentials than
single compartment reactors. The main feature
of these SSARs is that the mixing of various types
of sludge is prevented. These systems appear to be
particularly attractive for wastewater containing a
variety of soluble and insoluble and/or non-acidified
compounds3335.
The reported maximum organic loading rates
achieved at temperatures <15C are presented in
Table 3. However, the promising results depicted in
Table 3, which were mainly achieved in the past
20 years, are not sufficiently encouraging to implement
anaerobic wastewater treatment at full scale for the
treatment of cold wastewater (temperatures <18C).
In fact, only in the past ten years have higher rate
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Recycle

aValues

Wet gas meter

Effluent

Sludge bed

Influent

TRENDS in Biotechnology

Fig. 2. Schematic diagram of the UASB reactor system.

anaerobic wastewater treatment systems been

accepted, mainly for medium strength wastewater
under optimal mesophilic conditions.
The results of experiments into optimizing the
contact of sludge and wastewater in UASB reactors,
led to the development of an advanced reactor design,
namely the expanded granular sludge bed (EGSB;
Refs 27, 4042). The EGSB system uses exclusively
granular sludge (Fig. 4), whereas in anaerobic
fluidized bed (FB) or attached film expanded bed
(AFEB) reactors use inert carrier materials for
attachment of active biomass43. The upflow velocities
(vup) that can be applied in the EGSB system range
from 4 to 10 m h1. If necessary, these high vup values
can be achieved by applying effluent recycle and/or
using tall reactors.
The extraordinary high treatment efficiencies
achieved in EGSB reactors are more surprising in
view of the very low influent VFA concentrations of
0.50.8 g COD dm3 applied (Table 3; Ref. 16). The
VFA concentrations that prevail in the reactor
medium are in the range of 0.03 to 0.04 g COD dm3,
which indicates an extremely high substrate
affinity of the sludge at 10C. The estimated values
for the half saturation constant (Km) in the
MichaelisMenten equation for acetate, propionate
and butyrate amounted to 0.04, 0.01 and
0.14 g COD dm3, respectively44. The results
demonstrate the importance of adequate hydraulic
mixing in anaerobic systems for lowering the
apparent Km (Refs 38,42).
When applying a two-stage EGSB reactor
system, three to five times higher OLRs could be
accommodated at 90% CODvfa removal efficiency
than reported before for psychrophilic AnWT
(Table 3; Ref. 18). The enhancement of the
biodegradation process in a properly designed and

Fig. 3. Temperature
dependency of the
methane production rate
of mesophilic anaerobic
processes, after Henzen
and Harremoes29. White
diamond, Ref. 66; white
circle, Ref. 67; black
triangle, Ref. 68; black
circle, Ref. 8; black square,
Ref. 19; black diamond,
Ref. 69; cross, Ref. 70.
(Reproduced, with
permission, from Ref. 29)

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Activities at abscissa temperature

activity at 35 oC (%)

366

TRENDS in Biotechnology Vol.19 No.9 September 2001

130
110
90
70
60
50
40
30
20

10
0

10

20
30
40
Temperature (oC)

50

TRENDS in Biotechnology

operated staged reactor system can be attributed to

the development of a relatively balanced microecosystem in the sludge, in the various separate
reactor compartments. As a result of these
measures, the extent of product inhibition (e.g. of
hydrogen in the conversion of propionate) can be
reduced to a minimum. Consequently, in two-module
reactors, a compound such as propionate will be
degraded mainly in the second module, in which the
concentration of hydrogen and acetate will be
relatively low and consequently the conditions for
propionate degradation will be optimal. It is
becoming a self-regulating process; the consequence
of moduling is the development of sludge with an
exceptional high specific acetogenic and
methanogenic activity in the second module, leading
to a substantial increase in the organic loading
potentials of the system. A long-term experiment
conducted with malting wastewater in pilot-scale

two-stage EGSB reactor systems confirmed the

feasibility of the EGSB reactor design concept for
the treatment of cold low-strength complex
wastewater such as from malting process38.
There are still some important bottlenecks that
need to be eliminated, such as the occurrence of
components present in an acidification of
non-acidified wastewater (e.g. malting wastewater
and the adequate removal of suspended solids).
When imposing non-acidified wastewater to the
treatment using a methanogenic reactor under
psychrophilic conditions, a serious problem is the
formation of a layer of acidifying sludge around the
granules present in the EGSB or UASB reactors.
This layer formation can lead to gas entrapment
inside the granules, which initiates flotation of these
granules38,39. Under psychrophilic conditions,
acidifiers have a very high yield [0.22 g volatile
suspended solids (VSS)-COD g1 CODremoved; Ref. 20]
conditions compared with mesophilic conditions46
(Zoetemeyer, PhD thesis, Amsterdam University,
The Netherlands, 1982) owing to low starvation rate
of acidifiers at low temperature20. With respect to
insoluble matter, any degradation in psychrophilic
reactors cannot be expected because at low
temperatures (<15C) the hydrolysis rate of
suspended solids (SS) drops sharply and might even
approach zero for various types of solids47,48.
Nevertheless, it is remarkable that anaerobic
reactor systems were successfully operated at
exceptionally low temperatures (45C), which
confirms that stabile methanogenesis can be
achieved at such extreme low wastewater
temperatures18,37,39.
Development of granular psychrophilic biomass

The long-term operation of laboratory-scale and

pilot-scale EGSB reactors under psychrophilic
conditions resulted in the development of an active
methanogenic sludge that are well adapted to the low

Table 3. The anaerobic treatment of low strength wastewaters under psychrophilic conditions (<15C)
Reactor
type

Influent

Concentration
(g COD dm3)

OLR
(kg COD m3 d1)

Temperature
(C)

HRT
(h)

Efficiency
(%)

Refs

AAFEBa

Glucose

0.20.6

416

ASFb

Peptone

0.2h

0.64

10

16

4080

510

7.5

2735

UASBc

Sugar Vinasse

0.20.4

0.76.5

14

1.514

3265

EGSBd

VFA

2.6

2.0

27

12

32

50

27

UASBc

Beef consomm

1.47.0

210

10

16

4980

28

EGSBd

VFA

0.50.8

1012

1012

1.62.5

90

16

ASBRe

Dry milk

0.6

0.62.4

510

6585

37

EGSBf

Malting

0.21.8

312

1015

3.5

6778

38

EGSBf

VFA

0.50.9

512

48

24

90

18

UASBc

Wine Vinasse

1.25.2

0.37.3

411

1238

1592

39

UASBg

Wine Vinasse

1.15.4

0.85.5

410

1931

1680

39

aAnaerobic attached film expanded bed reactor. bAnaerobic submerged filter tank. cUpflow anaerobic sludge blanket reactor. dSingle stage
expanded granular sludge bed reactor. eAnaerobic sequencing batch reactor. fTwo stage expanded granular sludge bed reactor. gTwo stage
upflow anaerobic sludge blanket reactor. hg BOD dm3.

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Review

TRENDS in Biotechnology Vol.19 No.9 September 2001

Fig. 4. Schematic
diagram of the EGSB
reactor system. 1, feed;
2, feed distribution;
3, expanded sludge bed;
4, sieve drum; 5, gas liquid
separation; 6, effluent;
7, effluent recirculation;
8, biogas; 9, wet test gas
meter.

367

psychrophilic anaerobic wastewater treatment

requires the development of psychrophilic or
psychro-tolerant sub-populations, nor to what extent
mesophilic sludges can become psychro-tolerant.

8
9
5
6

Psychrophilic anaerobic sewage treatment

TRENDS in Biotechnology

temperature conditions16,37,39. Although they are very

likely to be present, the prevalence of psychrophilic
bacteria in the sludge have not yet been
demonstrated. Presumably, the psychrophilic
homologs in the sludge are not able to manifest in
these temperatureactivity profiles because of the
presence of many more mesophiles. In this context, it
should be understood that these mesophiles can still
grow prosperously under low temperature conditions
because the specific methanogenic activity of the
seed sludge improves significantly during the course
of the experiment at 10C (Refs 16,38). High
propionate degrading activities were also found at
5C (Ref. 18), which indicates satisfactory growth
and enrichment of these particular acetogens at
such low temperatures.
Furthermore, an observation of considerable
practical importance is that the methanogenic
sludge can preserve its achieved methanogenic
activity at low temperatures38. A six-month storage
period at 4C did not affect the methanogenic
capacity of the sludge. Apparently, the starvation
rate of methanogens grown at low temperatures is
extremely low. In practice, this means that
psychrophilically grown sludge will enable fast
start-up of new psychrophilic reactor systems.
It is not clear, as yet, whether high-rate
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The big challenge for engineers working in the field of

AnWT is to demonstrate the feasibility of AnWT for
sewage treatment, particularly in moderate climates.
Sewage belongs to the complex wastewater category
because (1) it contains a higher fraction particulate
COD, (2) the biodegradability of the various CODfractions is moderate, (3) it is a low strength
wastewater with varying concentrations and (4) its
temperature is often relatively low49.
Since the 1980s, comprehensive investigations
in this field have been conducted at several
laboratories worldwide. From the results obtained
(Table 4), it is clear that the anaerobic treatment of
domestic sewage might also present an attractive
and feasible option for moderate climates. However,
at the lower temperatures that prevail in the winter,
more sophisticated reactors are needed56,58,59
(Wang K., PhD thesis, Wageningen University, The
Netherlands, 1994). For the treatment of domestic
sewage, physical and biological processes are of
great importance.
The physical removal of particulate organics
owing to settling, adsorption and entrapment in the
sludge bed of the UASB is the first step in the
anaerobic treatment and conversion of domestic
sewage. The hydrolysis of retained particulate is, in
general, considered to be the rate-limiting step in the
overall digestion process and requires relatively long
retention times, depending on the applied process
temperature.
Degradation of the entrapped sewage particles in
anaerobic reactors is strongly affected under
psychrophilic conditions owing to a significant
decrease in the hydrolysis rate with declining
temperature60,61. Although a sludge retention time
(SRT) of 15 days is sufficient for providing hydrolysis
and methanogenesis at a process temperature of
25C (Ref. 61), an SRT of 75 days is required at a
process temperature of 15C (Ref. 60).. The longer
the required SRT, the shorter the minimum
hydraulic retention time (HRT) that can be applied60.
Hydrolysis of proteins and lipids strongly declines
with decreasing temperature, especially approaching
15C (ORourke J.T., PhD thesis, Stanford
University, CA, USA, 1968). Lipid hydrolysis will
only occur when full methanogenic conditions are
established61. The anaerobic hydrolysis rate of
particles has been shown to depend on the available
surface area and thus on the particle size62.
Researchers have proposed several surface-based
kinetic (SBK) models for the hydrolysis of
particulates6264.
Anaerobic treatment of domestic wastewater was
researched in both flocculent sludge and granular

368

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TRENDS in Biotechnology Vol.19 No.9 September 2001

Table 4. The anaerobic treatment of sewage under psychrophilic conditions (< 20C)
Reactor
type

Influent

Concentration
(g COD dm3)

OLR
Temperature
(kg COD m3 d1) (C)

HRT
(h)

Efficiency
(%)

Refs

UASBa

Row

0.3

0.6

820

12

67

50

UASBa

Row

0.4

1.2

1112

3050

51

AFb

Row

0.53

1.8

1315

3555

52

UASBa

Row

0.191.18

0.63.5

1220

78

3075

53

UASBa

Row

0.465

0.62

1218

18

73

54

FBc

Row

0.76

8.9

10

1.72.3

5385

55

EGSBd

Pre-settled

0.3

4.5

911

2.1

2048

48

UASB + EGSB

Row

0.320.51

1.62.5

813

4557

Wang K., 1994

AF + AHe

Row

0.460.53

0.91.1

13

12

70

56

UASB

Row

0.150.6

0.83.1

1325

4.7

6470

57

aUpflow

anaerobic sludge blanket reactor. bAnaerobic filter. cFluidized bed reactor. dSingle stage expanded granular sludge bed reactor.
filter + anaerobic hybrid reactor.

eAnaerobic

sludge bed reactors. Granular sludge will only

develop when pre-settled sewage is treated. When
treating raw domestic sewage at low temperatures
in the UASB reactors inoculated with granular
sludge, accumulation of SS occurs in the granular
sludge bed, which leads to deterioration of the
overall methanogenic activity and reactor
performance53,57. When using one-step flocculent
sludge bed reactors, the SRT should be long enough
to provide conditions for methanogenesis. At low
temperatures long SRTs are required resulting in
long HRTs (Ref. 60).
The particulate organic COD in domestic sewage
is, in general, divided into two fractions the
suspended-COD (>4.4 m) and the colloidal COD
(0.45 m < size <4.4 m). Several investigators
revealed that at low temperatures pre-removal of
CODSS is needed before anaerobic treatment in
granular bed reactors56,59,60. The first reactor
module in a staged reactor mainly serves for
removing major components of the CODSS.
Subsequent modules (e.g. consisting of either a
UASB-, EGSB- or AH-anaerobic hybrid reactor
systems) serve in the removal of soluble and/or
colloidal compounds. Application of a two-step
UASB-system, instead of a one-step system was
shown to improve the removal of particulate,
especially suspended, COD (Ref. 47). At
temperatures as low as 12C, however, still only
42% suspended COD could be removed in the first
step. Moreover, the removal of colloidal COD
(CODcol) was limited in the applied two-step UASB
plus EGSB system47. Because the biodegradability
of CODcol is high, the low removal of CODcol in
continuous anaerobic reactors at low temperature
should be attributed to the low entrapment and
sorption in the sludge bed65.
Elmitwalli and colleagues investigated the
removal of CODss and CODcol at low temperatures56.
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They discovered that removal could be enhanced by

the introduction of filter material in a two-step
process. An anaerobic filter (AF) reactor with vertical
sheets of reticulated polyurethane foam (RPF),
applied as a first step at an HRT of 4 h and a
temperature of 13C, showed a high removal
efficiency for CODss (82%) . Moreover, the
performance of the AF was more stable in comparison
to other tested systems, especially during rainy
periods. Because the AF is operated at short HRT and
low temperature, the hydrolysis, acidification and
methanogenesis is limited and, therefore, the excess
sludge produced needs post stabilization.
The treatment of pre-settled sewage instead of
raw sewage in a UASB and an AH reactor at 13C
significantly increased CODcol removal efficiency
for both reactors. The AH reactor is recommended
for the treatment of pre-treated sewage at low
temperatures because a significantly higher
removal of CODcol can be achieved. Based on the
above-mentioned results, it is concluded that the
most suitable configuration for the treatment of
domestic sewage at low temperatures is an AF
reactor followed by an AH reactor56. At the
application of a two-step AF plus AH system at an
HRT of 4 plus 8 h and 13C, high removal
efficiencies for all COD fractions are achieved.
The total COD (CODtot) removal amounts to 71%,
which is similar to that found in tropical countries
with constant high temperatures56. In the imposed
conditions, a removal of CODcol of 60% is achieved in
a two-step system but the removal efficiency of this
fraction could be increased by the addition of
polymers. Biomass and colloidal particles have a
negative charge65. Addition of a small amount of a
cationic polymer before the second step might
improve the removal of colloidal particles as a result
of the observed increase in the size of the colloidal
particles56 (Elmitwally T., PhD thesis, Wageningen

Review

TRENDS in Biotechnology Vol.19 No.9 September 2001

University, The Netherlands, 2000). Given that an

important part of pathogens is associated with
colloidal particles, an increased removal of latter
fraction will be accompanied with a higher removal
of pathogens56.
Conclusions

Especially when using staged EGSB-reactors,

psychrophilic AnWT is feasible and is now open for
full scale applications to low strength, mainly or
partially acidified wastewater, including various
types of industrial wastewater and pre-settled
domestic sewage. Also temperatures below 10C
can probably be applied.
The prospect of anaerobic sewage treatment
using UASB or EGSB reactors seems particularly
promising for regions in which sewage temperatures
do not drop below 15C. Such conditions might
prevail in subtropical regions, such as in the
Mediterranean countries. In addition, in moderate
climates the AnWT concept represents an attractive
solution, although this might require a change in
sanitation paradigms. Moreover, of particular
interest with respect to the treatment of sewage is
the observation of the formation of a granular sludge
of a relatively high specific activity on settled sewage.
When domestic sewage is treated anaerobically
under low temperature conditions, a two-step system
combined with an additional sludge digester is
required. The application of the combined AF or AH
system results in high removal efficiencies. When
nutrient removal is obligatory, a different system
configuration in which the second step is operated as
a denitrifying reactor can be chosen.
For sewage treatment at low temperatures
(<15C) and also when the temperature fluctuates
between 15C (winter) and 25C (summer), a
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