Wat. Res. Vol. 34, No. 15, pp.

3867±3875, 2000
7 2000 Elsevier Science Ltd. All rights reserved
Printed in Great Britain
PII: S0043-1354(00)00136-6 0043-1354/00/$ - see front matter

www.elsevier.com/locate/watres

TREATMENT OF DILUTE WASTEWATER USING AN

ANAEROBIC BAFFLED REACTOR: EFFECT OF LOW
TEMPERATURE
ALETTE A. M. LANGENHOFF$M and DAVID C. STUCKEY*M
Department of Chemical Engineering, Imperial College, Prince Consort Road, London SW7 2BY, UK

(First received 1 June 1999; accepted in revised form 1 January 2000)

AbstractÐThe performance of an anaerobic ba‚ed reactor (ABR, 10 l and eight compartments)

treating a dilute wastewater (500 mg COD/l) was studied. The reactor was started with a hydraulic
retention time (HRT) of 80 h at 358C, and this was steadily reduced to 10 h eventually; all HRTs
tested resulted in more than 80% COD removal. The temperature was then reduced to 208C, which
gave a removal eciency of 70%, and ®nally to 108C, which resulted in only 60% COD removal.
Residence time distribution studies were carried out to monitor hydrodynamic ¯ow characteristics and
dead space after each decrease in temperature. Not much di€erence in mixing or dead space was
observed under the di€erent conditions tested. The average dead space was 25±30%, and the ¯ow
pattern within the reactors showed an intermediate behaviour between plug-¯ow and ideally mixed.
Finally, anaerobic bioassays were used to evaluate the activity of the biomass. The biomass retained its
initial speci®c acetogenic and methanogenic activity throughout the compartments of the reactor, and
hardly any selection for psychrophilic bacteria had taken place. 7 2000 Elsevier Science Ltd. All
rights reserved

Key wordsÐanaerobic digestion, low strength feed, soluble, low temperature, anaerobic ba‚ed reactor

INTRODUCTION temperatures lower than 358C, heating is required

The treatment of both domestic and industrial
wastewaters is carried out primarily using biological
methods due to their lower costs compared to
chemical methods. Great advances have been made
over the last 20 years in anaerobic reactor design,
and in understanding the complex microbial pro-
cesses that occur in anaerobic digestion (Speece,
1996). These advances have enabled reactors to
maintain a high solids retention time (SRT 20±100
days), while keeping the hydraulic retention time
(HRT) to a minimum (1.3±20 h), and have enabled
a variety of dilute wastes, such as sewage, to be
treated economically. The successful treatment of
low strength (soluble and colloidal) wastewaters at
low retention times in an anaerobic ba‚ed reactor
(ABR) at mesophilic temperatures has been
described elsewhere (Langenho€ et al., 2000). Most
full-scale treatment systems operate under mesophi-
lic temperatures (30±358C). However, since a num-
ber of wastewaters, including domestic sewage, have
*Author to whom all correspondence should be addressed.
Tel.: +44-171-5945591; fax: +44-171-5945629; e-mail:
d.stuckey@ic.ac.uk
$
Present address: TNO-MEP, PO Box 342, 7300 AH
Apeldoorn, The Netherlands.
3867
thus substantially increasing the costs of treating
this waste. Therefore, anaerobic treatment systems
able to operate at low temperatures (10±208C) o€er
the potential of substantially lower treatment costs
for low temperature wastewaters, and could poten-
tially increase the range of anaerobic treatment to
domestic sewage.
Reducing the operating temperature a€ects the
performance of an anaerobic reactor as biological
processes are a€ected by temperature. Overall,
lower substrate concentrations and reduced tem-
peratures result in a deterioration of process per-
formance compared to the case of high substrate
concentration and high temperatures (Nachaiyasit
and Stuckey, 1997). Several studies have examined
the performance of anaerobic treatment plants at
low temperatures; in most cases mesophilic sludge
was used to start-up the reactors, after which time
the temperature was slowly decreased. Tempera-
tures as low as 58C have been used, with removal
eciencies ranging from 0 to 90% in anaerobic ®l-
ters (Cullimore and Viraraghavan, 1994), ¯uidised
bed reactors (Sanz and Fdz-Polanco, 1990), anaero-
bic sequencing batch reactors (ASBR) (Ndon and
Dague, 1997), and up¯ow anaerobic sludge bed
reactors (UASB) (Kettunen and Rintala, 1998). Stu-
3868 Alette A. M. Langenho€ and David C. Stuckey
dies that looked at the microbiology in more detail
at low temperatures concluded that although the
total amount of bacteria in the reactor only slowly
reduced when the temperature was decreased from
30 to 158C, the rate of acetogenesis dropped signi®-
cantly, and some time was needed before it regained
its original activity (Cha et al., 1997).
The ABR performs well in treating medium
(1±10 g COD/l) and high strength (>10 g COD/
l) soluble wastewater at mesophilic temperatures,
with COD removal eciencies of more than 95%
(Barber and Stuckey, 1999). The advantage of
such a compartmentalised reactor is the partial
separation of acidogenic and methanogenic pro-
cesses (Hasouna and Stuckey, 2000), its stability
to hydraulic shocks, and its ability to separate
the liquid (HRT) and solid (SRT) retention times
(Grobicki and Stuckey, 1991). Furthermore, when
treating wastewater at low temperature, the com-
partmentalisation might enhance the hydrolysis of
less easily degradable substrates in the front of
the reactor due to its low pH. Nevertheless, at
low temperatures the enhanced production of sol-
uble microbial products (SMP) may increase the
e‚uent COD signi®cantly (Schiener et al., 1998).
These SMP can be de®ned as compounds of mi-
crobial origin which result from substrate metab-
olism and biomass decay, and have been found
to account for the majority of soluble organic
material in the e‚uent from biological treatment
processes (Schiener et al., 1998). Due to the com-
partmentalised structure of the ABR, the pro-
duction and degradation of SMP can be followed
more closely than in a mixed system like a CSTR
or UASB, and enable greater insights to be
obtained about the role of SMPs in low tempera-
ture treatment.
Hence, the aim of this research was to evaluate
the performance of anaerobic ba‚ed reactors in
treating dilute wastewaters (0500 mg COD/l) at
di€erent temperatures (35, 20 and 108C), and to
determine if the mixing characteristics in the reactor
depended on temperature. In addition, batch bioas-
says were performed to gain more insight into the
microbial selection processes that were occurring in
the reactor, and the in¯uence of temperature on mi-
crobial activity.
MATERIALS AND METHODS
Experimental set-up
The studies were performed in a 10 l ABR made of
clear acrylic plastic. The reactor contained vertical hanging
and standing ba‚es which divided it into eight identical
compartments, and was operated and seeded as previously
described (Langenho€ et al., 2000). The feed consisted of
pasteurised semi-skimmed milk diluted with tap water to a
®nal concentration of 500 mg COD/l. The reactor was in-
itially maintained at 358C in a water bath, and the data
obtained at this temperature provided an internal control
in order to compare performance at lower temperatures.
Starting at an HRT of 80 h, the HRT was gradually
decreased to 6 h, but due to deteriorating performance it
was returned back to 10 h HRT. After reaching steady-
state at 358C and 10 h HRT, the temperature was
decreased to 208C on day 175, and after 260 days to 108C.
Hydrodynamic ¯ow characteristics
Residence time distribution (RTD) studies were carried
out to analyse the hydrodynamics of the reactor by adding
a pulse of inert tracer (lithium) to the reactor. A 10 ml sol-
ution of lithium chloride (10 g/l-Sigma) was used, and the
concentration of lithium in the e‚uent was measured by
atomic adsorption spectroscopy (Perkin Elmer 1100 B) for
at least three HRTs after the addition of the pulse. The
recovery of the lithium was more than 90% in all exper-
iments.
Batch assays
The biodegradability of SMP in the e‚uent of the
ABR, of glucose and lactose by anaerobic sludge, and the
temperature dependence of the acetogenic and methano-
genic activity of the biomass inside the compartments of
the ABR were tested using bioassays. The bioassays were
based on the method of Owen et al. (1979), and performed
in 30-ml bottles ®lled with 10 ml of anaerobic medium
(Langenho€ et al., 1999). The gas phase used in the bottles
was N2/CO2 (70/30%). The bottles were inoculated with
approximately 0.1 g volatile suspended solids (VSS) of bio-
mass, and incubated at 358C. All experiments were per-
formed in duplicate, and controls without substrate were
also run.
To test the biodegradability of the various molecular
weight fractions of the SMP, the e‚uent of the ABR at
208C was ®rst separated into di€erent molecular weight
fractions using ultra®ltration (Barker et al., 1999). Sludge
from an anaerobic digester (Mogden, UK) was used as the
seed for the anaerobic assays, and the assays were incu-
bated for 2 months. These assays had a coecient of vari-
ation of 22%. An aerobic bacterial mixed culture
(Polyseed, Camlab, UK) was used for the aerobic assays
and incubated for 2 days. The degradation was calculated
as the decrease in COD concentration over time in the
assays.
The degradation of glucose and lactose (3 g COD/l) was
carried out using an unacclimatised seed from a sewage
sludge digester (Mogden, UK). The assays were incubated
at di€erent temperatures, and a decrease in substrate con-
centration over time was used to compare the activities of
the biomass on the two substrates. These two sugars were
chosen to determine if lactose metabolism changed in a
di€erent way to glucose with temperature.
The temperature dependence of the acetogenic and
methanogenic activity of the biomass was tested in similar
batches, and was inoculated with 1 ml of sludge (approxi-
mately 0.2 g VSS; APHA, 1992) withdrawn from the bot-
tom of each compartment of the ABR. Biomass was taken
on day 154 (358C), day 233 (208C), day 386 (108C), and
was incubated at 35, 20 and 108C, respectively. The
methanogenic activity in the batches was determined
by adding a VFA mixture (acetate:propionate:buty-
rate=1:1.5:1.8 by mass, resulting in 1 g COD/l of each
fatty acid) to result in a total COD in the assays of 3 g/l,
and measuring the methane produced. Methane pro-
duction was recorded daily using a wetted glass syringe to
measure the total gas produced, and measuring its compo-
sition by GC-TCD. The speci®c methane production after
10 days of incubation was calculated and used to compare
the di€erent incubations. To determine the acetogenic ac-
tivity of the biomass in the ABR, glucose and lactose were
used as substrates in di€erent bottles at a concentration of
3 g COD/l. 50 mM of bromoethanesulphonic acid (BESA)
was added to each bottle to inhibit methanogenic activity
 Low temperature anaerobic treatment
(Grbic-Galic, 1986) with the result that intermediate pro-
ducts accumulated. Microbial activity was calculated by
measuring the accumulation of VFAs. All experiments
were performed in duplicate.
Sampling and analyses
Several parameters (pH, VFAs, COD, VFA, lithium,
CH4, and CO2) were measured routinely in the feed, the
e‚uent, the eight compartments of the reactors, and the
batch experiments to characterise the behaviour of the
reactors and the activities in the bioassays, and have been
described previously (Langenho€ et al., 1999).
RESULTS AND DISCUSSION
Reactor performance
High removal eciencies were achieved when
treating the dilute wastewater at 358C at HRTs
down to 10 h (Fig. 1). Each decrease in HRT was
followed by a temporary increase in COD concen-
tration in the e‚uent, but the reactor quickly recov-
ered its removal eciency. The average COD
removal was more than 90% for an HRT of 10 h,
and the remaining COD in the e‚uent consisted
mainly of VFAs, indicating that the incoming COD
was almost completely degraded to biomass, CH4
and CO2. If a complex mixture like milk is
degraded, the concentration of VFAs in the e‚uent
will always be greater than zero due to the Ks
values of the methanogens, and limitations due to
mass transfer.
The Arrhenius equation (Levenspiel, 1972), pre-
dicts that a decrease in temperature results in a
decrease in reaction rate, and hence the biological
activity (Qo) will decrease by a factor of about 3
with a temperature drop of 158C. As can be seen in
Fig. 1, changing the temperature from 35 to 208C
resulted in a decrease in removal eciency of only
25% (95 4 70%), not as much as predicted by the
Arrhenius equation. This shows that the reactor
contained more than enough active biomass to treat
the incoming wastes, and this was not degrading
the incoming COD at its maximum rate at 358C.
Furthermore, another drop in temperature from 20
Fig. 1. Chemical oxygen demand, VFA concentrations,
and COD removal eciencies in the e‚uent of the reactor.
The changes in HRT and temperature are given at the top
of the ®gure.
3869
to 108C resulted in only a 10% decrease in removal
capacity, again highlighting the surplus active bio-
mass in the reactor. Most mesophilic reactors are
usually thought to possess a metabolic overcapacity
(i.e. they are capable of catabolising considerably
more feed at maximum growth rates than is being
fed), as is also shown in our reactor, making them
more stable to shocks, e.g. a decrease in tempera-
ture.
If VFAs accumulate to any signi®cant degree in
the reactor (>150 mg/l), this is the ®rst indication
that the system is not operating at optimal con-
ditions. At 208C the reactor started to show the
®rst signs of decreased reactor performance when
VFAs (100 ppm, mainly acetate) were detected in
the e‚uent, and this contributed to the majority of
COD found in the e‚uent (75%). The production
of fatty acids at low temperatures occurs at a rela-
tive higher rate than methanogenesis, and this
results in the accumulation of intermediate fatty
acids (Nozhevnikova et al., 1994). For 3 months
after the temperature change to 108C, the COD
concentration in the e‚uent increased, and the
VFA data showed that the majority of the COD in
the e‚uent consisted of VFAs; this also occurred in
the temperature drop from 35 to 208C. It appears
that poor contact between biomass and substrate,
due to lower gas production and mixing at lower
temperatures, was responsible for this decrease in
reactor performance (Nachaiyasit and Stuckey,
1997). However, after 60 days of operation at 108C
the total VFA concentration in the reactor e‚uent
started to decrease from 180 mg COD/l to only
50 mg COD/l. As the amount of COD in the e‚u-
ent remained fairly constant at 200 mg COD/l, the
contribution of the VFAs to COD in the e‚uent
decreased from 90 to only 25%.
The other 75% of COD in the e‚uent could not
be characterised/analysed, and was therefore
denoted as SMP; this increased from 15 mg/l as
COD at 358C to 40 mg/l at 208C and 150 mg/l at
108C. A comparable increase in COD in the e‚uent
from an ABR due to a decrease in temperature has
also been noted (Schiener et al., 1998). Only a few
papers quantify the VFA contribution to COD in
the e‚uent at low temperatures, and this contri-
bution varies from 80 to 99% (Kettunen and Rin-
tala, 1998; Ndon and Dague, 1997), which was not
the case in this study. All the milk was metabolised
in the reactor (data not shown), but whether the
SMP in the e‚uent at 108C resulted from substrate
metabolism or biomass decay could not determined.
However, the reactor e‚uent at 108C initially con-
tained most of the COD in the form of VFAs (day
260±320), indicating that the biomass in the reactor
was able to acidify the substrate, and that the pro-
duction of intermediate products other than VFAs
was minimised at 108C. After 320 days of operation
with a low strength wastewater, 60 days of which
were at 108C, the SMP concentration in the e‚uent
3870 Alette A. M. Langenho€ and David C. Stuckey
Fig. 2. Residence time distribution (C-curve) of the ABR
at di€erent temperatures.
increased. One plausible explanation for this was
that this change was due to an increased production
of extracellular polymers (ECPs), and this change
occurred at the expense of VFA production. It is
known that both conditions, i.e. low substrate con-
centrations and low temperatures, can enhance the
production of ECP (Troy, 1979). In this case more
ECPs were produced at low temperatures and
slowly released from the bacterial cell wall, and it
appears there was a substantial time lag, possibly
due to selection. These microbial products could
have been the cause of an increased SMP concen-
tration in the e‚uent at low temperature. A few
psychrophilic bacteria have been described in the
literature, and most of them produce slime (Noz-
hevnikova et al., 1994), most likely contributing to
the SMP in the e‚uent.
Residence time distributions
Residence time distribution (RTD) studies were
carried out at di€erent temperatures to determine
whether the rate of gas production and viscosity
a€ected the hydrodynamics of the ABR. The nor-
malised concentration of the lithium tracer in the
e‚uent was plotted against the normalised time in
a C-curve, and the results are shown in Fig. 2. The
data from these graphs were analysed with a two-
phase dispersion model (Grobicki and Stuckey,
1992; Levenspiel, 1972), and the results are shown
in Table 1. The volume of dead space was relatively
constant at the temperatures used, even though less
gas was produced at low temperature due to the
lower COD removals.
Using the calculated dispersion numbers it can be
concluded that the ¯ow pattern within the reactor
Table 1. Results of the residence time distribution (RTD) studies and SMP production in the ABR with an HRT of 10 h at di€erent tem-
HRT Dead space (%) Variance d 2
358C 25.4 0.10
208C 30.4 0.15
108C 27.1 0.21
a
D/ml is the dispersion number, and is the reciprocal of the Peclet number, Pe. Variance, d 2, is the variance of the C curve (dimensionless
contraction vs time). N is the number of completely mixed tanks (CSTRs) in series.
Fig. 3. Speci®c methane-producing activity (after day 10)
of the biomass in the ABR at 358C (a) and 108C (b), and
incubated at 35, 20 and 108C.
was intermediate between plug-¯ow and perfectly
mixed under all the conditions tested. Any deviation
from ideality could have been caused by channelling
of the liquid through the biomass bed, or by the
creation of dead spaces in the reactor.
E‚uent biodegradability
The biodegradability of the COD in the e‚uent
(VFA and SMP) at day 233 (ABR at 208C) was
tested in both aerobic and anaerobic batch assays
(Table 2). The e‚uent was divided into di€erent
molecular fractions using membrane separation to
distinguish between the degradability of these frac-
tions. Under aerobic conditions all the fractions
showed good degradability, except for the low mol-
ecular weight fraction of less than 1000 Dalton,
however, the COD of this fraction was so low as to
make this result meaningless. A di€erent result was
found under anaerobic conditions with only the
lowest molecular weight fraction being degradable,
indicating that the high molecular weight com-
pounds were very hard to degrade under anaerobic
conditions. As the e‚uent contained mainly high
peraturesa
D/ml N Gas production (l/d) Average COD removal
0.054 9.6 3.0 95%
0.083 6.5 2.2 70%
0.083 4.7 1.9 60%
 Low temperature anaerobic treatment
molecular weight fractions, this shows that the e‚u-
ent at low temperature had a low biodegradability
under anaerobic conditions. Based on these data it
appears that to increase the biodegradation and
performance of the ABR, the introduction of an
aerobic stage in one of the last compartments might
prove useful.
Methanogenic activity of the biomass
The batch assays carried out with the 358C seed
biomass from the ABR showed an irregular pattern
of bacterial activity at an incubation temperature of
358C for the various compartments, with lower ac-
tivities at 20 and 108C (Fig. 3a), as predicted by the
Arrhenius equation. High methanogenic activities
were not only found at the inlet of the reactor
(compartment 2 and 3), but also at the end of the
reactor at 358C. Given this trend, it may be that the
sample from compartment 4 was not representative.
Incubations at 208C also showed higher methano-
genic activities at the beginning of the reactor than
towards the end. This implies that there was no
enrichment for fermentative and acetogenic bacteria
at the beginning of the reactor, or methanogenic
bacteria towards the end of the reactor. Current
understanding of the ABR suggests that it is a com-
partmentalised reactor that has the potential to
partially or totally separate the acidogenic and
methanogenic phases, but apparently this is not
true for ABRs treating low strength wastewater.
The initial seed sludge still appears to be present
throughout the reactor, without any selection of the
speci®c groups of microorganisms in the separate
compartments after 5 months of operation. It
appears from other work (Hasouna and Stuckey,
2000) that due to the low gas production with low
strength wastewaters there is little washout of bio-
mass from the reactor, and hence little physical
selection pressure.
The inoculum from the 108C reactor (Fig. 3b)
showed di€erent activities in a number of compart-
ments compared to the inoculum at 358C. When
the 108C sludge was incubated at 358C, an extre-
mely high methanogenic activity was found for the
®rst compartment, while in the third compartment
it was also high. Activities in the other compart-
ments were generally lower than the inoculum from
the ABR at 358C. Various groups of methanogens
Table 2. Aerobic and anaerobic biodegradability of molecular weight fractions of the ABR e‚uent at 208C
MW fraction (Dalton) Concentration in e‚uent (mg COD/l)
MW > 300k 48
MW < 300k 5 95
MW > 100k 70
MW < 100k 8 100
MW > 10k 76
MW < 10k 14
MW > 1k 76
MW < 1k 3 17
Total sample 90
3871
must have been involved in the production of
methane, each with a di€erent temperature sensi-
tivity, since di€erent activities were found with the
biomass from the compartments at di€erent tem-
peratures. Whereas the 208C incubation with the
358C inoculum (Fig. 3a) showed a higher activity at
the inlet of the reactor, the 208C incubation with
the 108C inoculum (Fig. 3b) showed a higher ac-
tivity towards the outlet of the reactor. This was
probably due to the slowdown of fermentation at
lower temperatures resulting in more VFAs (which
are the substrates for the methanogenic bacteria)
being formed towards the middle and end of the
reactor. Another explanation for the selection of
the methanogens towards the end of the reactor is
that this biomass was taken from the reactor at day
386, which meant that the reactor had been oper-
ated twice as long before sampling compared to the
biomass at 358C. A selection of the speci®c groups
of microorganisms of the initial seed sludge over
the separate compartments could have taken place
after such a long operation time.
Since the activity of the 108C inoculum showed a
high activity when incubated at its original tempera-
ture of 358C (Fig. 3b), this data shows that the
dominant populations were still mesophilic even
after 4 months of operation under psychrophilic
conditions. The increased activity in compartment 1
might have been due to the fact that not all the bio-
mass at 358C in the ABR was active, since the seed
was taken from a long retention time (20 days) sew-
age sludge digester. When operating the reactor at
108C, more biomass needed to be active to degrade
the incoming substrate at the low temperature; this
will not necessarily be expressed in the speci®c ac-
tivity since the VSS did not increase much.
Acetogenic activity of the biomass
A more comprehensive study was performed on
the acetogenic activity of the biomass obtained
from the reactor. Both glucose and lactose were
used as substrates, and gave comparable degra-
dation rates, because the sludge in the ABR was
adapted to milk which contains lactose in contrast
to unadapted sludge gave di€erent results (data not
shown). The methanogens in the assays were inhib-
ited by BESA, and VFAs accumulated in the bot-
tles. The VFAs produced by seed from the eight
Aerobic biodegradability (%) Anaerobic biodegradability (%)
86 4
12
96 8
0
74 7
64 0
84 1
33
89 N.D.
 3872
Alette A. M. Langenho€ and David C. Stuckey

Fig. 4. VFA pro®les of batches using biomass taken from each compartment of the ABR at 358C, and incubated at 358C (BESA inhibited).
 Low temperature anaerobic treatment

Fig. 5. VFA pro®les of the batches incubated with biomass and glucose as a substrate from compartment 3 of the ABR. Conditions are (1): 358C inoculum, incubated at 358C, (2): 208C
inoculum, incubated at 358C, (3): 108C inoculum, incubated at 358C, (4): 358C inoculum, incubated at 208C, (5): 208C inoculum, incubated at 208C, (6): 108C inoculum, incubated at 208C,
(7): 358C inoculum, incubated at 108C, (8): 208C inoculum, incubated at 108C, (9): 108C inoculum, incubated at 108C.
3873
3874 Alette A. M. Langenho€ and David C. Stuckey
di€erent compartments of the ABR at 358C (taken
at day 154), and incubated at 358C, can be seen in
Fig. 4. As can be seen, the activity throughout the
reactor was fairly constant except for compartment
7, as was found for the methanogenic activity
(Fig. 3), again indicating that enrichment for
speci®c groups of bacteria in the compartments had
not yet taken place, even after 5 months of incu-
bation. However, the production of acetate, propio-
nate, and butyrate was high in compartment 7, in
comparison to the other compartments, indicating a
high acetogenic activity in compartment 7, and we
do not have an explanation for this at present.
Acetate, the main precursor for methane production
(Schink, 1988), was formed in high concentrations
(1500 ppm), with smaller amounts of propionate in
a ratio of 4 to 1. Formate was often detected in the
®rst few days, but disappeared rapidly. Higher fatty
acids, and ethanol and lactate were not detected in
the bioassays, indicating a balanced degradation
process, and more than 90% of the glucose was
recovered in the VFAs formed, except for compart-
ment 7.
The e€ect of incubation temperature, and adap-
tation to temperature, of the acetogenic bacteria
can be seen in batch assays with biomass adapted
to di€erent temperatures, and incubated at di€erent
temperatures (Fig. 5). Compartment 3 was used as
an example, but the other compartments gave com-
parable pro®les. Incubations at 358C with di€erent
temperature inocula gave similar results; the for-
mation of acetate and propionate in a ratio of 1 to
3 or 4 (Fig. 5.1-3). This again shows that the domi-
nant populations were still mesophilic after 5
months of operation under psychrophilic con-
ditions, as the biomass collected at 108C (Fig. 5.3)
had virtually the same speci®c activity and product
spectrum as the biomass collected at 358C (Fig. 5.1).
The incubations with low temperature sludge incu-
bated at low temperatures (Fig. 5.5-6-8-9) showed
either more propionate production than acetate, or
equal amounts. It seems that the propionate
degrading bacteria (syntrophs) are more a€ected by
temperature than the acetate degrading bacteria
(methanogens), and hence propionate accumulates
in the system. Eventually the propionate concen-
tration decreases, but it takes more time compared
to the incubations at higher temperatures. Further-
more, the accumulation of propionate in an anaero-
bic reactor is an indication of unstable anaerobic
process. This can be con®rmed by the results in our
reactor at 108C, which did not perform as well as at
358C.
The formation and degradation of butyrate did
not seem to be a€ected by low temperatures, as
no butyrate accumulated in the system. When lac-
tose was added as a substrate to the batch assays,
it was degraded to intermediates in a few days
(results not shown), such as propionate, butyrate,
long chain fatty acids, and these compounds were
further oxidised by acetogenic bacteria to acetate,
CO2, hydrogen and formate. Since the long chain
fatty acids and butyrate were not detected in our
batch incubations, these degradation reactions were
less in¯uenced by the temperature drop. This has
been demonstrated in EGSB reactors as well; in ex-
periments with biomass from the reactor operated
at low temperature and incubated at low tempera-
ture, the degradation of acetate and propionate
were most sensitive to low temperatures. A 18C
drop in temperature lead to a substantial decrease
in acetate and propionate removal, whereas the
degradation of butyrate was more stable, and less
a€ected by a change in temperature (Rebac et al.,
1995).
CONCLUSIONS
. It has been shown that low strength wastewaters
can be treated successfully at 358C (195%
removal), while at 208C the removal eciency
was adequate (70%). However, at 108C COD
removals dropped to 60%. The performance of
the ABR at high ¯owrates (10 h HRT) and low
temperature shows that the treatment of low
strength wastewater is possible at low tempera-
tures, providing the opportunity to decrease the
costs of domestic wastewater treatment systems.
However, an aerobic polishing step would be use-
ful to degrade the high molecular weight fraction
of SMPs produced in the reactor at 108C.
. The mixing characteristics of the reactor can be
modelled by a two-phase dispersion model, and
the eight compartments behave like eight separate
CSTRs in series. Decreases in temperature lead-
ing to increased viscosity and decreased gas pro-
duction did not seem to in¯uence the amount of
dead space in the reactors, and hence this par-
ameter is controlled by other more complex vari-
ables.
. The biomass in the ABR retained its initial
speci®c methanogenic and acetogenic activity at
mesophilic temperature, even after incubation for
many months at psychrophilic temperatures. This
shows that the mesophilic biomass in the ABR
was not too sensitive to temperature shocks.
Hardly any selection or enrichment for psychro-
philic bacteria had taken place, but the mesophi-
lic bacteria initially present were metabolising at
lower rates at 25 and 108C. Mesophilic inocula
incubated at lower temperatures in batch bioas-
says produced more propionate than acetate due
to a greater reduction in the rate of the syntrophs
compared to the methanogens at low tempera-
tures.
. Selection for speci®c groups of fermentative,
acetogenic and methanogenic bacteria over the
di€erent compartments did not take place when
treating a low strength wastewater.
 Low temperature anaerobic treatment
AcknowledgementsÐThis work was supported by a grant
from the Engineering and Physical Sciences Research
Council (EPSRC), UK.
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