Você está na página 1de 9

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



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 thus substantially increasing the costs of treating
wastewaters is carried out primarily using biological this waste. Therefore, anaerobic treatment systems
methods due to their lower costs compared to able to operate at low temperatures (10±208C) o€er
chemical methods. Great advances have been made the potential of substantially lower treatment costs
over the last 20 years in anaerobic reactor design, for low temperature wastewaters, and could poten-
and in understanding the complex microbial pro- tially increase the range of anaerobic treatment to
cesses that occur in anaerobic digestion (Speece, domestic sewage.
1996). These advances have enabled reactors to Reducing the operating temperature a€ects the
maintain a high solids retention time (SRT 20±100 performance of an anaerobic reactor as biological
days), while keeping the hydraulic retention time processes are a€ected by temperature. Overall,
(HRT) to a minimum (1.3±20 h), and have enabled lower substrate concentrations and reduced tem-
a variety of dilute wastes, such as sewage, to be peratures result in a deterioration of process per-
treated economically. The successful treatment of formance compared to the case of high substrate
low strength (soluble and colloidal) wastewaters at concentration and high temperatures (Nachaiyasit
low retention times in an anaerobic ba‚ed reactor and Stuckey, 1997). Several studies have examined
(ABR) at mesophilic temperatures has been the performance of anaerobic treatment plants at
described elsewhere (Langenho€ et al., 2000). Most low temperatures; in most cases mesophilic sludge
full-scale treatment systems operate under mesophi- was used to start-up the reactors, after which time
lic temperatures (30±358C). However, since a num- the temperature was slowly decreased. Tempera-
ber of wastewaters, including domestic sewage, have 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
*Author to whom all correspondence should be addressed. bed reactors (Sanz and Fdz-Polanco, 1990), anaero-
Tel.: +44-171-5945591; fax: +44-171-5945629; e-mail:
bic sequencing batch reactors (ASBR) (Ndon and
Present address: TNO-MEP, PO Box 342, 7300 AH Dague, 1997), and up¯ow anaerobic sludge bed
Apeldoorn, The Netherlands. 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 Starting at an HRT of 80 h, the HRT was gradually
at low temperatures concluded that although the decreased to 6 h, but due to deteriorating performance it
was returned back to 10 h HRT. After reaching steady-
total amount of bacteria in the reactor only slowly state at 358C and 10 h HRT, the temperature was
reduced when the temperature was decreased from decreased to 208C on day 175, and after 260 days to 108C.
30 to 158C, the rate of acetogenesis dropped signi®-
cantly, and some time was needed before it regained Hydrodynamic ¯ow characteristics
its original activity (Cha et al., 1997). Residence time distribution (RTD) studies were carried
The ABR performs well in treating medium out to analyse the hydrodynamics of the reactor by adding
(1±10 g COD/l) and high strength (>10 g COD/ 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
l) soluble wastewater at mesophilic temperatures, concentration of lithium in the e‚uent was measured by
with COD removal eciencies of more than 95% atomic adsorption spectroscopy (Perkin Elmer 1100 B) for
(Barber and Stuckey, 1999). The advantage of at least three HRTs after the addition of the pulse. The
such a compartmentalised reactor is the partial recovery of the lithium was more than 90% in all exper-
separation of acidogenic and methanogenic pro- iments.
cesses (Hasouna and Stuckey, 2000), its stability
Batch assays
to hydraulic shocks, and its ability to separate
the liquid (HRT) and solid (SRT) retention times The biodegradability of SMP in the e‚uent of the
ABR, of glucose and lactose by anaerobic sludge, and the
(Grobicki and Stuckey, 1991). Furthermore, when temperature dependence of the acetogenic and methano-
treating wastewater at low temperature, the com- genic activity of the biomass inside the compartments of
partmentalisation might enhance the hydrolysis of the ABR were tested using bioassays. The bioassays were
less easily degradable substrates in the front of based on the method of Owen et al. (1979), and performed
in 30-ml bottles ®lled with 10 ml of anaerobic medium
the reactor due to its low pH. Nevertheless, at (Langenho€ et al., 1999). The gas phase used in the bottles
low temperatures the enhanced production of sol- was N2/CO2 (70/30%). The bottles were inoculated with
uble microbial products (SMP) may increase the approximately 0.1 g volatile suspended solids (VSS) of bio-
e‚uent COD signi®cantly (Schiener et al., 1998). mass, and incubated at 358C. All experiments were per-
These SMP can be de®ned as compounds of mi- formed in duplicate, and controls without substrate were
also run.
crobial origin which result from substrate metab- To test the biodegradability of the various molecular
olism and biomass decay, and have been found weight fractions of the SMP, the e‚uent of the ABR at
to account for the majority of soluble organic 208C was ®rst separated into di€erent molecular weight
material in the e‚uent from biological treatment fractions using ultra®ltration (Barker et al., 1999). Sludge
from an anaerobic digester (Mogden, UK) was used as the
processes (Schiener et al., 1998). Due to the com- seed for the anaerobic assays, and the assays were incu-
partmentalised structure of the ABR, the pro- bated for 2 months. These assays had a coecient of vari-
duction and degradation of SMP can be followed ation of 22%. An aerobic bacterial mixed culture
more closely than in a mixed system like a CSTR (Polyseed, Camlab, UK) was used for the aerobic assays
or UASB, and enable greater insights to be and incubated for 2 days. The degradation was calculated
as the decrease in COD concentration over time in the
obtained about the role of SMPs in low tempera- assays.
ture treatment. The degradation of glucose and lactose (3 g COD/l) was
Hence, the aim of this research was to evaluate carried out using an unacclimatised seed from a sewage
the performance of anaerobic ba‚ed reactors in sludge digester (Mogden, UK). The assays were incubated
at di€erent temperatures, and a decrease in substrate con-
treating dilute wastewaters (0500 mg COD/l) at centration over time was used to compare the activities of
di€erent temperatures (35, 20 and 108C), and to the biomass on the two substrates. These two sugars were
determine if the mixing characteristics in the reactor chosen to determine if lactose metabolism changed in a
depended on temperature. In addition, batch bioas- di€erent way to glucose with temperature.
says were performed to gain more insight into the The temperature dependence of the acetogenic and
methanogenic activity of the biomass was tested in similar
microbial selection processes that were occurring in batches, and was inoculated with 1 ml of sludge (approxi-
the reactor, and the in¯uence of temperature on mi- mately 0.2 g VSS; APHA, 1992) withdrawn from the bot-
crobial activity. 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
MATERIALS AND METHODS by adding a VFA mixture (acetate:propionate:buty-
rate=1:1.5:1.8 by mass, resulting in 1 g COD/l of each
Experimental set-up fatty acid) to result in a total COD in the assays of 3 g/l,
The studies were performed in a 10 l ABR made of and measuring the methane produced. Methane pro-
clear acrylic plastic. The reactor contained vertical hanging duction was recorded daily using a wetted glass syringe to
and standing ba‚es which divided it into eight identical measure the total gas produced, and measuring its compo-
compartments, and was operated and seeded as previously sition by GC-TCD. The speci®c methane production after
described (Langenho€ et al., 2000). The feed consisted of 10 days of incubation was calculated and used to compare
pasteurised semi-skimmed milk diluted with tap water to a the di€erent incubations. To determine the acetogenic ac-
®nal concentration of 500 mg COD/l. The reactor was in- tivity of the biomass in the ABR, glucose and lactose were
itially maintained at 358C in a water bath, and the data used as substrates in di€erent bottles at a concentration of
obtained at this temperature provided an internal control 3 g COD/l. 50 mM of bromoethanesulphonic acid (BESA)
in order to compare performance at lower temperatures. was added to each bottle to inhibit methanogenic activity
Low temperature anaerobic treatment 3869

(Grbic-Galic, 1986) with the result that intermediate pro- to 108C resulted in only a 10% decrease in removal
ducts accumulated. Microbial activity was calculated by
capacity, again highlighting the surplus active bio-
measuring the accumulation of VFAs. All experiments
were performed in duplicate. mass in the reactor. Most mesophilic reactors are
usually thought to possess a metabolic overcapacity
Sampling and analyses (i.e. they are capable of catabolising considerably
Several parameters (pH, VFAs, COD, VFA, lithium, more feed at maximum growth rates than is being
CH4, and CO2) were measured routinely in the feed, the fed), as is also shown in our reactor, making them
e‚uent, the eight compartments of the reactors, and the
batch experiments to characterise the behaviour of the more stable to shocks, e.g. a decrease in tempera-
reactors and the activities in the bioassays, and have been ture.
described previously (Langenho€ et al., 1999). 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
Reactor performance VFAs (100 ppm, mainly acetate) were detected in
High removal eciencies were achieved when the e‚uent, and this contributed to the majority of
treating the dilute wastewater at 358C at HRTs COD found in the e‚uent (75%). The production
down to 10 h (Fig. 1). Each decrease in HRT was of fatty acids at low temperatures occurs at a rela-
followed by a temporary increase in COD concen- tive higher rate than methanogenesis, and this
tration in the e‚uent, but the reactor quickly recov- results in the accumulation of intermediate fatty
ered its removal eciency. The average COD acids (Nozhevnikova et al., 1994). For 3 months
removal was more than 90% for an HRT of 10 h, after the temperature change to 108C, the COD
and the remaining COD in the e‚uent consisted concentration in the e‚uent increased, and the
mainly of VFAs, indicating that the incoming COD VFA data showed that the majority of the COD in
was almost completely degraded to biomass, CH4 the e‚uent consisted of VFAs; this also occurred in
and CO2. If a complex mixture like milk is the temperature drop from 35 to 208C. It appears
degraded, the concentration of VFAs in the e‚uent that poor contact between biomass and substrate,
will always be greater than zero due to the Ks due to lower gas production and mixing at lower
values of the methanogens, and limitations due to temperatures, was responsible for this decrease in
mass transfer. reactor performance (Nachaiyasit and Stuckey,
The Arrhenius equation (Levenspiel, 1972), pre- 1997). However, after 60 days of operation at 108C
dicts that a decrease in temperature results in a the total VFA concentration in the reactor e‚uent
decrease in reaction rate, and hence the biological started to decrease from 180 mg COD/l to only
activity (Qo) will decrease by a factor of about 3 50 mg COD/l. As the amount of COD in the e‚u-
with a temperature drop of 158C. As can be seen in ent remained fairly constant at 200 mg COD/l, the
Fig. 1, changing the temperature from 35 to 208C contribution of the VFAs to COD in the e‚uent
resulted in a decrease in removal eciency of only decreased from 90 to only 25%.
25% (95 4 70%), not as much as predicted by the The other 75% of COD in the e‚uent could not
Arrhenius equation. This shows that the reactor be characterised/analysed, and was therefore
contained more than enough active biomass to treat denoted as SMP; this increased from 15 mg/l as
the incoming wastes, and this was not degrading COD at 358C to 40 mg/l at 208C and 150 mg/l at
the incoming COD at its maximum rate at 358C. 108C. A comparable increase in COD in the e‚uent
Furthermore, another drop in temperature from 20 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
Fig. 1. Chemical oxygen demand, VFA concentrations,
and COD removal eciencies in the e‚uent of the reactor. was minimised at 108C. After 320 days of operation
The changes in HRT and temperature are given at the top with a low strength wastewater, 60 days of which
of the ®gure. 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 Fig. 3. Speci®c methane-producing activity (after day 10)
appears there was a substantial time lag, possibly of the biomass in the ABR at 358C (a) and 108C (b), and
due to selection. These microbial products could incubated at 35, 20 and 108C.
have been the cause of an increased SMP concen-
tration in the e‚uent at low temperature. A few was intermediate between plug-¯ow and perfectly
psychrophilic bacteria have been described in the mixed under all the conditions tested. Any deviation
literature, and most of them produce slime (Noz- from ideality could have been caused by channelling
hevnikova et al., 1994), most likely contributing to of the liquid through the biomass bed, or by the
the SMP in the e‚uent. creation of dead spaces in the reactor.

Residence time distributions E‚uent biodegradability

Residence time distribution (RTD) studies were The biodegradability of the COD in the e‚uent
carried out at di€erent temperatures to determine (VFA and SMP) at day 233 (ABR at 208C) was
whether the rate of gas production and viscosity tested in both aerobic and anaerobic batch assays
a€ected the hydrodynamics of the ABR. The nor- (Table 2). The e‚uent was divided into di€erent
malised concentration of the lithium tracer in the molecular fractions using membrane separation to
e‚uent was plotted against the normalised time in distinguish between the degradability of these frac-
a C-curve, and the results are shown in Fig. 2. The tions. Under aerobic conditions all the fractions
data from these graphs were analysed with a two- showed good degradability, except for the low mol-
phase dispersion model (Grobicki and Stuckey, ecular weight fraction of less than 1000 Dalton,
1992; Levenspiel, 1972), and the results are shown however, the COD of this fraction was so low as to
in Table 1. The volume of dead space was relatively make this result meaningless. A di€erent result was
constant at the temperatures used, even though less found under anaerobic conditions with only the
gas was produced at low temperature due to the lowest molecular weight fraction being degradable,
lower COD removals. indicating that the high molecular weight com-
Using the calculated dispersion numbers it can be pounds were very hard to degrade under anaerobic
concluded that the ¯ow pattern within the reactor conditions. As the e‚uent contained mainly high

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 D/ml N Gas production (l/d) Average COD removal

358C 25.4 0.10 0.054 9.6 3.0 95%

208C 30.4 0.15 0.083 6.5 2.2 70%
108C 27.1 0.21 0.083 4.7 1.9 60%

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.
Low temperature anaerobic treatment 3871

molecular weight fractions, this shows that the e‚u- must have been involved in the production of
ent at low temperature had a low biodegradability methane, each with a di€erent temperature sensi-
under anaerobic conditions. Based on these data it tivity, since di€erent activities were found with the
appears that to increase the biodegradation and biomass from the compartments at di€erent tem-
performance of the ABR, the introduction of an peratures. Whereas the 208C incubation with the
aerobic stage in one of the last compartments might 358C inoculum (Fig. 3a) showed a higher activity at
prove useful. the inlet of the reactor, the 208C incubation with
the 108C inoculum (Fig. 3b) showed a higher ac-
Methanogenic activity of the biomass tivity towards the outlet of the reactor. This was
The batch assays carried out with the 358C seed probably due to the slowdown of fermentation at
biomass from the ABR showed an irregular pattern lower temperatures resulting in more VFAs (which
of bacterial activity at an incubation temperature of are the substrates for the methanogenic bacteria)
358C for the various compartments, with lower ac- being formed towards the middle and end of the
tivities at 20 and 108C (Fig. 3a), as predicted by the reactor. Another explanation for the selection of
Arrhenius equation. High methanogenic activities the methanogens towards the end of the reactor is
were not only found at the inlet of the reactor that this biomass was taken from the reactor at day
(compartment 2 and 3), but also at the end of the 386, which meant that the reactor had been oper-
reactor at 358C. Given this trend, it may be that the ated twice as long before sampling compared to the
sample from compartment 4 was not representative. biomass at 358C. A selection of the speci®c groups
Incubations at 208C also showed higher methano- of microorganisms of the initial seed sludge over
genic activities at the beginning of the reactor than the separate compartments could have taken place
towards the end. This implies that there was no after such a long operation time.
enrichment for fermentative and acetogenic bacteria Since the activity of the 108C inoculum showed a
at the beginning of the reactor, or methanogenic high activity when incubated at its original tempera-
bacteria towards the end of the reactor. Current ture of 358C (Fig. 3b), this data shows that the
understanding of the ABR suggests that it is a com- dominant populations were still mesophilic even
partmentalised reactor that has the potential to after 4 months of operation under psychrophilic
partially or totally separate the acidogenic and conditions. The increased activity in compartment 1
methanogenic phases, but apparently this is not might have been due to the fact that not all the bio-
true for ABRs treating low strength wastewater. mass at 358C in the ABR was active, since the seed
The initial seed sludge still appears to be present was taken from a long retention time (20 days) sew-
throughout the reactor, without any selection of the age sludge digester. When operating the reactor at
speci®c groups of microorganisms in the separate 108C, more biomass needed to be active to degrade
compartments after 5 months of operation. It the incoming substrate at the low temperature; this
appears from other work (Hasouna and Stuckey, will not necessarily be expressed in the speci®c ac-
2000) that due to the low gas production with low tivity since the VSS did not increase much.
strength wastewaters there is little washout of bio-
mass from the reactor, and hence little physical Acetogenic activity of the biomass
selection pressure. A more comprehensive study was performed on
The inoculum from the 108C reactor (Fig. 3b) the acetogenic activity of the biomass obtained
showed di€erent activities in a number of compart- from the reactor. Both glucose and lactose were
ments compared to the inoculum at 358C. When used as substrates, and gave comparable degra-
the 108C sludge was incubated at 358C, an extre- dation rates, because the sludge in the ABR was
mely high methanogenic activity was found for the adapted to milk which contains lactose in contrast
®rst compartment, while in the third compartment to unadapted sludge gave di€erent results (data not
it was also high. Activities in the other compart- shown). The methanogens in the assays were inhib-
ments were generally lower than the inoculum from ited by BESA, and VFAs accumulated in the bot-
the ABR at 358C. Various groups of methanogens tles. The VFAs produced by seed from the eight

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) Aerobic biodegradability (%) Anaerobic biodegradability (%)

MW > 300k 48 86 4
MW < 300k 5 95 12
MW > 100k 70 96 8
MW < 100k 8 100 0
MW > 10k 76 74 7
MW < 10k 14 64 0
MW > 1k 76 84 1
MW < 1k 3 17 33
Total sample 90 89 N.D.
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.
3874 Alette A. M. Langenho€ and David C. Stuckey

di€erent compartments of the ABR at 358C (taken further oxidised by acetogenic bacteria to acetate,
at day 154), and incubated at 358C, can be seen in CO2, hydrogen and formate. Since the long chain
Fig. 4. As can be seen, the activity throughout the fatty acids and butyrate were not detected in our
reactor was fairly constant except for compartment batch incubations, these degradation reactions were
7, as was found for the methanogenic activity less in¯uenced by the temperature drop. This has
(Fig. 3), again indicating that enrichment for been demonstrated in EGSB reactors as well; in ex-
speci®c groups of bacteria in the compartments had periments with biomass from the reactor operated
not yet taken place, even after 5 months of incu- at low temperature and incubated at low tempera-
bation. However, the production of acetate, propio- ture, the degradation of acetate and propionate
nate, and butyrate was high in compartment 7, in were most sensitive to low temperatures. A 18C
comparison to the other compartments, indicating a drop in temperature lead to a substantial decrease
high acetogenic activity in compartment 7, and we in acetate and propionate removal, whereas the
do not have an explanation for this at present. degradation of butyrate was more stable, and less
Acetate, the main precursor for methane production a€ected by a change in temperature (Rebac et al.,
(Schink, 1988), was formed in high concentrations 1995).
(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 CONCLUSIONS
acids, and ethanol and lactate were not detected in
the bioassays, indicating a balanced degradation . It has been shown that low strength wastewaters
process, and more than 90% of the glucose was can be treated successfully at 358C (195%
recovered in the VFAs formed, except for compart- removal), while at 208C the removal eciency
ment 7. was adequate (70%). However, at 108C COD
The e€ect of incubation temperature, and adap- removals dropped to 60%. The performance of
tation to temperature, of the acetogenic bacteria the ABR at high ¯owrates (10 h HRT) and low
can be seen in batch assays with biomass adapted temperature shows that the treatment of low
to di€erent temperatures, and incubated at di€erent strength wastewater is possible at low tempera-
temperatures (Fig. 5). Compartment 3 was used as tures, providing the opportunity to decrease the
an example, but the other compartments gave com- costs of domestic wastewater treatment systems.
parable pro®les. Incubations at 358C with di€erent However, an aerobic polishing step would be use-
temperature inocula gave similar results; the for- ful to degrade the high molecular weight fraction
mation of acetate and propionate in a ratio of 1 to of SMPs produced in the reactor at 108C.
3 or 4 (Fig. 5.1-3). This again shows that the domi- . The mixing characteristics of the reactor can be
nant populations were still mesophilic after 5 modelled by a two-phase dispersion model, and
months of operation under psychrophilic con- the eight compartments behave like eight separate
ditions, as the biomass collected at 108C (Fig. 5.3) CSTRs in series. Decreases in temperature lead-
had virtually the same speci®c activity and product ing to increased viscosity and decreased gas pro-
spectrum as the biomass collected at 358C (Fig. 5.1). duction did not seem to in¯uence the amount of
The incubations with low temperature sludge incu- dead space in the reactors, and hence this par-
bated at low temperatures (Fig. 5.5-6-8-9) showed ameter is controlled by other more complex vari-
either more propionate production than acetate, or ables.
equal amounts. It seems that the propionate . The biomass in the ABR retained its initial
degrading bacteria (syntrophs) are more a€ected by speci®c methanogenic and acetogenic activity at
temperature than the acetate degrading bacteria mesophilic temperature, even after incubation for
(methanogens), and hence propionate accumulates many months at psychrophilic temperatures. This
in the system. Eventually the propionate concen- shows that the mesophilic biomass in the ABR
tration decreases, but it takes more time compared was not too sensitive to temperature shocks.
to the incubations at higher temperatures. Further- Hardly any selection or enrichment for psychro-
more, the accumulation of propionate in an anaero- philic bacteria had taken place, but the mesophi-
bic reactor is an indication of unstable anaerobic lic bacteria initially present were metabolising at
process. This can be con®rmed by the results in our lower rates at 25 and 108C. Mesophilic inocula
reactor at 108C, which did not perform as well as at incubated at lower temperatures in batch bioas-
358C. says produced more propionate than acetate due
The formation and degradation of butyrate did to a greater reduction in the rate of the syntrophs
not seem to be a€ected by low temperatures, as compared to the methanogens at low tempera-
no butyrate accumulated in the system. When lac- tures.
tose was added as a substrate to the batch assays, . Selection for speci®c groups of fermentative,
it was degraded to intermediates in a few days acetogenic and methanogenic bacteria over the
(results not shown), such as propionate, butyrate, di€erent compartments did not take place when
long chain fatty acids, and these compounds were treating a low strength wastewater.
Low temperature anaerobic treatment 3875

AcknowledgementsÐThis work was supported by a grant (2000) The anaerobic treatment of dilute soluble and
from the Engineering and Physical Sciences Research colloidal wastewater using an anaerobic ba‚ed reactor;
Council (EPSRC), UK. in¯uence of hydraulic retention time. Wat. Res. 34,
Levenspiel O. (1972) Chemical Reaction Engineering, 2nd
REFERENCES ed. John Wiley & Sons, New York.
Nachaiyasit S. and Stuckey D. C. (1997) E€ect of low
APHA (1992) Standard Methods for the Examination of temperatures on the performance of an anaerobic
Water and Wastewater, 18th ed. American Public Health ba‚ed reactor (ABR). J. Chem. Technol. Biotechnol. 69,
Association, Washington. 276±284.
Barber W. P. and Stuckey D. C. (1999) The use of the an-
Ndon U. J. and Dague R. R. (1997) E€ect of temperature
aerobic ba‚ed reactor (ABR) for wastewater treatment;
and hydraulic retention time on anaerobic sequencing
a review. Wat. Res. 33, 1559±1578.
batch reactor treatment of low-strength wastewater.
Barker D. J., Mannucchi G. A., Salvi S. M. L. and
Wat. Res. 31, 2455±2466.
Stuckey D. C. (1999) Characterisation of soluble re-
Nozhevnikova A. N., Kotsyurbenko O. R. and Simankova
sidual chemical oxygen demand (COD) in anaerobic
M. V. (1994) Acetogenesis at low temperature. In Aceto-
wastewater treatment e‚uents. Wat. Res. 33, 2499±
genesis, ed. H. L. Drake, pp. 416±444. Chapman &
Cha G. C., Chung H. K. and Chung J. C. (1997) Suppres- Hall, New York.
sion of acidogenic activities due to rapid temperature Owen W. F., Stuckey D. C., Healy J. B. J., Young L. Y.
drop in anaerobic digestion. Biotechnol. Letters 19, 461± and McCarty P. L. (1979) Bioassay for monitoring bio-
464. chemical methane potential and anaerobic toxicity. Wat.
Cullimore D. R. and Viraraghavan T. (1994) Microbial Res. 13, 485±492.
aspects of anaerobic ®lter treatment of septic tank e‚u- Rebac S., Ruskova J., Gerbens S., van Lier J. B., Stams
ent at low temperature. Environ. Technol. 15, 163±173. A. J. M. and Lettinga G. (1995) High-rate anaerobic
Grbic-Galic D. (1986) Anaerobic production and trans- treatment of wastewater under psychrophilic conditions.
formation of aromatic hydrocarbons and substituted J. Ferm. Bioeng. 80, 499±506.
phenols by ferulic acid-degrading BESA-inhibited Sanz I. and Fdz-Polanco F. (1990) Low temperature treat-
methanogenic consortia. FEMS Microbiol. Ecol. 38, ment of municipal sewage in anaerobic ¯uidized bed
161±169. reactors. Wat. Res. 24, 463±469.
Grobicki A. and Stuckey D. C. (1991) Performance of the Schiener P., Nachaiyasit S. and Stuckey D. C. (1998) Pro-
anaerobic ba‚ed reactor under steady-state and shock duction of soluble microbial products (SMP) in an an-
loading conditions. Biotechnol. Bioeng. 37, 344±355. aerobic ba‚ed reactor: composition, biodegradability,
Grobicki A. and Stuckey D. C. (1992) Hydrodynamic and the e€ect of process parameters. Environ. Technol.
characteristics of the anaerobic ba‚ed reactor. Wat. 19, 391±399.
Res. 26, 371±378. Schink B. (1988) Principles and limits of anaerobic degra-
Hasouna S. E. and Stuckey D. C. (2000) The e€ect of or- dation: environmental and technological aspects. In Bi-
ganic loading rate on the separation of bacterial trophic ology of Anaerobic Microorganisms, ed. A. J. B.
groups in an anaerobic ba‚ed reactor. Wat. Res. sub- Zehnder, pp. 771±846. John Wiley & Sons, New York.
mitted. Speece R. E. (1996) Anaerobic Biotechnology for Industrial
Kettunen R. H. and Rintala J. A. (1998) Performance of Wastewater. Archae Press, Nashville, Tennessee.
an on-site UASB-reactor treating leachate at low tem- Troy II F. A. (1979) The chemistry and biosynthesis of
perature. Wat. Res. 32, 537±546. selected bacterial capsular polymers. Annu. Rev. Micro-
Langenho€ A. A. M., Intrachandra N. and Stuckey D. C. biol. 33, 519±560.