Treatment of sewage by a UASB reactor under moderate to low

temperature conditions
Shigeki Uemura
a,
*
, Hideki Harada
b
a
Department of Civil and Environmental Engineering, Kisarazu National College of Technology, 2-11-1 Kiyomidai-higashi, Kisarazu, Chiba 292-0041,
Japan
b
Department of Environmental Systems Engineering, Nagaoka University of Technology, 1603-1 Kamitomioka, Nagaoka, Niigata 940-2188, Japan
Received 19 December 1998; received in revised form 5 July 1999; accepted 25 July 1999
Abstract
The feasibility of sewage treatment by an upow anaerobic sludge blanket (UASB) reactor was studied using actual sewage at a
xed hydraulic retention time (HRT) of 4.7 h, and at temperatures in the range of 2513C, for six months. The average total COD
removals and solid COD removals achieved were 70% and 80%, respectively. Total COD removal rate depended on inuent
strength, especially solid COD concentration, rather than operational temperature. Particulate organic matter in the inuent was
eectively removed by entrapment in the sludge bed. The hydrolysis rate of the entrapped organics was signicantly aected by
temperature, that is, 58% of the entrapped particulate organics was liqueed at 25C, decreasing to 33% at 13C. The methanogenic
activity of the retained sludge decreased eventually to 410% of the seed granular sludge which had been formed on carbohydrate
wastewater. Microscopic observation of the morphology of the sludge revealed that even though the sludge remained in granular
form as a whole, the granules tended to be partly broken in a state of autolysis. 1999 Elsevier Science Ltd. All rights reserved.
Keywords: UASB reactor; Anaerobic sewage treatment; Methane fermentation; Liquefaction; Methanogenic activity
1. Introduction
In the last two decades, anaerobic treatment tech-
nology has developed remarkably for the treatment of
specic industrial wastewaters, mainly consisting of
soluble and easily degradable organic substances in high
concentrations. This great success in the development of
anaerobic wastewater technology can be attributed to
the introduction of innovative bioreactors, i.e., anaero-
bic xed bed, anaerobic uidized bed, and UASB reac-
tors. Even though current anaerobic technology seems
to have reached maturity in the treatment of certain
types of industrial wastewater, its application to waste-
waters which contain high concentrations of particulate
organics (lipids, proteins), and inhibitory organic and/or
inorganic substances, is still restricted.
One of the next challenges for anaerobic technology
is the extension of its applicability to lower-strength
wastewaters, i.e., sewage and domestic wastewater (van
der Last and Lettinga, 1992). The direct treatment of
sewage by an anaerobic process is undoubtedly an at-
tractive and appropriate option, especially for develop-
ing countries, since it requires low energy for operation
and low initial investment, and also provides lower
sludge production and easier maintenance than con-
ventional aerobic processes.
In many developing countries, which are confronting
rapidly increasing water pollution problems, this tech-
nology becomes even more favorable and promising,
since the warm climate in these countries makes the
process more eecient (Agrawal et al., 1997; Alaerts
et al., 1993; Tare et al., 1997).
On this basis, a long-term study was conducted to
assess the feasibility of sewage treatment by a UASB
reactor using actual sewage. The main objectives of this
study were (1) to investigate the eect of temperature on
the process performance under relatively moderate
temperature conditions, (2) to assess the mass balance
on COD at dierent temperatures, and (3) to quantify
the microbial activity of the sludge retained in the re-
actor in terms of methanogenic activity, ATP content
and coenzyme F
420
content.
Bioresource Technology 72 (2000) 275282
*
Corresponding author. Tel.: +81-438-30-4152; fax: +81-438-98-
5717.
E-mail address: cuemura@minato.kisarazu.ac.jp (S. Uemura).
0960-8524/00/$ - see front matter 1999 Elsevier Science Ltd. All rights reserved.
PII: S 0 9 6 0 - 8 5 2 4 ( 9 9 ) 0 0 1 1 8 - 2
2. Methods
2.1. UASB reactor
The schematic diagram of the experimental apparatus
is shown in Fig. 1. The reactor consisted of a poly-
acrylamide column of 2.0 m in height and 10.0 cm in
diameter, on the top of which a gassolidsliquid sepa-
rator (GSS) was installed. The total volume of the re-
actor was 21.5 l (15.7-l column and 5.8-l GSS). In order
to assist de-gassication of the rising sludge before en-
tering into the GSS, three inverted cones with 6 mm
perforations over 16% of their surface area were in-
stalled in the upper portion of the column. A gas re-
circulation pump was used for 30 s per 15 min to break
the scum. In addition, a scum breaker was incorporated
within the GSS and was operated at 120 rpm for 30 s per
15 min.
2.2. Wastewater
The sewage from our university was employed as
inuent. This wastewater derives mainly from restau-
rants, laboratories and dormitories, and does not con-
tain inhibitory substances from laboratories. To
minimize any time uctuation in concentration, the
sewage was collected daily, at a dened time when COD
reached its peak value, from a sewer pipe just prior to its
municipal sewer junction. The raw wastewater was l-
tered through a 60-mesh screen basket to remove large
suspended solids.
2.3. Seed sludge
Granular sludge was used as an inoculum sludge. An
amount of 390 g mixed liquor suspended solids (MLSS)
with 91% of mixed liquor volatile suspended solids
(MLVSS) was seeded to the reactor. This inoculum was
obtained from a lab-scale reactor (210-l capacity) in our
laboratory, in which the sludge had been cultivated on a
mixture of starch and sucrose at a loading rate of 11 kg
COD m
3
d
1
(inuent concentration: 3000 mg l
1
,
HRT: 6.5 h) for a period of three months after estab-
lishment of a complete granular sludge bed.
2.4. Experimental procedure
The UASB reactor was operated by feeding the
sewage inuent for a period of six months. The HRT
was xed at 4.7 h throughout the experiment. The re-
actor was placed in a temperature control chamber. The
continuous experiment was conducted at dierent tem-
peratures, reducing from 25C to 13C in 3C steps. The
reactor was operated at 25C for the rst 57 days, for the
following 36 days at 22C, and for 31 days at 19C, 29
days at 16C, and 25 days at 13C. The temperature
decrease from 25C to 13C for a duration of 6 months
might not be long enough for adaptation of anaerobic
microorganisms to low temperature conditions. How-
ever, one of the purposes of this study was to estimate
the process performance of a UASB reactor when
treating sewage under relatively moderate temperature
conditions. Therefore, in order to simulate the temper-
ature decrease in countries with moderate climate, we
decided on the above experimental duration.
2.5. Analytical procedure
The volatile fatty acids (VFAs) and gas composition
were determined by the method described previously
(Harada et al., 1994). The measurement of COD, sul-
fate, MLSS, MLVSS and sludge volume index (SVI)
were done following the Japanese Standard Methods for
Sewage and Wastewater (Japan Sewage Works Associ-
ation, 1997). Preparation of COD samples was con-
Fig. 1. Schematics of experimental apparatus: (1) substrate reservoir;
(2) stirrer; (3) feeding pump; (4) inuent; (5) sampling port; (6) per-
forated corn; (7) scum breaker; (8) euent; (9) GSS; (10) gas recircu-
lation line; (11) air pump; (12) scum trap; (13) gas trap; (14) wet test
gas meter.
276 S. Uemura, H. Harada / Bioresource Technology 72 (2000) 275282
ducted as described in previous reports (Agrawal et al.,
1997; Harada et al., 1994).
The ATP pool of the sludge was determined accord-
ing to the method of Petterson et al. (1970). Anaero-
bically dispersed sludge under an O
2
-free N
2
gas
atmosphere was resuspended in 0.025 M tris buer (pH
7.5) and kept at 100C for 10 min for extraction. The
extract was analyzed by the uorescence assay method
using an ATP photometer (Aminco, J4-7441 with reac-
tion chamber). The extraction of coenzyme F
420
was
done according to the procedure described by Dolng
and Mulder (1985). Scanning electron microscope
(SEM) observation was done as in our previous report
(Uemura and Harada, 1993). The methanogenic activity
of the sludge was determined by means of a serum-vial
test, using the method reported previously (Harada
et al., 1994). The C/N ratio of sludge samples was de-
termined by a CN analyzer (Yanako MT-500, Yanako,
Japan).
3. Results
3.1. Reactor performance
Time courses of COD in the inuent and in the ef-
uent are given in Fig. 2. The inuent concentration
uctuated greatly, in the range of 115595 mg l
1
for
total COD (mean: 312 mg l
1
, STD: 73.2). Soluble COD
ranged from 63 to 245 mg l
1
(mean: 114 mg l
1
, STD:
30.4). Accordingly, particulate COD comprised ap-
proximately 60% of the total COD. The volumetric
COD loading rate was 1.6 kg m
3
d
1
(range 0.73.0).
The euent total COD was in the range of 49174 mg
l
1
(mean: 89 mg l
1
, STD: 25.8). The soluble COD in
the euent lay in the range of 35104 mg l
1
(mean: 50
mg l
1
, STD: 11.8). The euent water qualities in terms
of total COD and soluble COD were relatively stable
throughout the experiment and seemed to be not sig-
nicantly aected by operational temperature. The
COD removal was 69.4% on average (6085%) based on
the inuent total and the euent total, and 84% (66
92%) based on the inuent total and the euent soluble
(Fig. 2). It should be noted that VFAs in the euent
were rarely detected (detectable limit was 1 ppm)
throughout the experiment (data not shown). The solid
COD in the euent ranged from 7.5 to 82 mg l
1
(mean
38 mg l
1
). The average pH values of the inuent and
euent were 6.86 (6.27.4) and 6.98 (6.227.5), respec-
tively, maintaining nearly the optimum level for the
methane fermentation process.
3.2. Methane production
Fig. 3 represents the time course for total methane
production and the recovery of the biogas. Hereafter,
net methane production refers to the total CH
4
pro-
duction per day, that is, the sum of the amount of CH
4
collected and that escaped in dissolved form in the ef-
uent. The dissolved CH
4
content was calculated by
Henry's Law. A considerably large fraction of CH
4
gas
escaped in dissolved form in the euent.
Fig. 4 indicates the relationship between total COD
removal and solid COD removal for the entire temper-
ature range. Although the plots for each temperature are
scattered over both the abscissa and ordinate, solid
COD removal was linearly correlated with total COD
removal. This indicates that the total COD removal was
signicantly aected by the inuent solid COD, rather
than by temperature.
Table 1 represents the relationship between the COD
removal rate and CH
4
production rate for each tem-
perature. The ratios of the amount of CH
4
to COD
removal for each temperature, determined by linear
regression, are given in the table. In the table, the total
COD removal rate was based on inuent total COD
and euent total COD. The CH
4
production rate was
calculated as the sum of the amount of CH
4
collected
and that discharged in the euent. The amount of CH
4
recovered from the reactor was estimated to be less
than 0.23 Nl g
1
COD removed for the whole temper-
ature range. Even taking into account the amount of
Fig. 2. Time course of inuent COD, euent COD and COD removal eciency. Symbols: (h) COD removal eciency based on inuent total COD
and euent soluble COD; (s) inuent total COD; (d) inuent soluble COD; (M) euent total COD; (.) euent solid COD.
S. Uemura, H. Harada / Bioresource Technology 72 (2000) 275282 277
CH
4
discharged in dissolved form, the ratio was still in
the range of 0.160.27 Nl g
1
COD removed. Consid-
ering that the theoretical ratio is 0.35 Nl g
1
COD
removed, this suggested that a part of the inuent
solid COD remained within the reactor without being
liqueed.
3.3. COD balance
Table 2 presents the inuence of temperature on the
solid COD removal (%), soluble COD removal (%),
total COD removal (%), solid COD accumulated in the
reactor (%), liqueed fraction of solid COD accumu-
lated in the reactor (%), methane recovery (%) and
sludge retention time (SRT) in 13C and 25C phases. In
the table, the dierence between the amount of input
(solid COD+soluble COD) and that of output (solid
COD+soluble COD+COD as recovered CH
4
+COD
as dissolved CH
4
discharged with the euent +COD
used for sulfate reduction) can be regarded as the
amount of solid COD accumulated in the reactor (re-
ferred to as accumulated COD). The liqueed fraction
Fig. 4. Relationship between total COD removal and solid COD removal. Total COD removedinuent total CODeuent total COD. Solid
COD removedinuent solid CODeuent solid COD. Symbols: (s) 25C; (M) 22C; (h) 19C; () 16C; (d) 13C.
Fig. 3. Time course of methane production. Symbols: (s) recovered CH
4
as biogas; (d) recovered CH
4
+dissolved CH
4
.
Table 1
The ratio of methane production rate to COD removal rate in each temperature
Temperature (C) 25 22 19 16 13
CH
4
production rate/COD removal rate 0.26 0.27 0.18 0.16 0.16
278 S. Uemura, H. Harada / Bioresource Technology 72 (2000) 275282
was calculated as (1 accumulated COD/(inuent solid
CODeuent solid COD)). With regard to the result
at 19C, the eect of the strength of the inuent solid
COD masked the eect of temperature on these COD
reduction parameters. As a whole, even though the solid
and total COD removals were also slightly aected by
temperature reduction, the liqueed fraction signi-
cantly decreased as temperature decreased. Soluble
COD removal was eventually independent of the tem-
perature, hence the level of the euent soluble COD
remained almost constant throughout the experiment.
These results implied that the removal of solid COD was
due mainly to entrapment within the sludge bed.
Methane recovery was dened as the ratio of amount
of methane recovered as biogas to that of total CH
4
generated. As expected from Fig. 3, methane recovery
exhibited a tendency to decrease considerably as tem-
perature decreased (in this case 3560%).
3.4. Substrate removal pattern
Fig. 5 shows the substrate removal patterns along the
height of the reactor for the nal days of operation at
25C and 13C. Soluble COD in the inuent was 120
and 70 mg l
1
, respectively, for 25C and 13C. This
increased to 250265 mg l
1
(for 25C) and 270280 mg
l
1
(for 13C) at a height of 1020 cm from the bottom
due to the liquefaction of entrapped solid organics, and
then decreased along the reactor height due to the fur-
ther progress of methanization. Similarly, the total
VFAs accumulated reached 2530 mg COD l
1
(for
25C) and 100150 mg COD l
1
(for 13C) at a height of
1020 cm, then converted rapidly into methane and
disappeared completely at the outlet. Within the bed,
only acetate and propionate but no higher VFAs accu-
mulated for all the temperatures. The existence of higher
concentrations of soluble COD can be attributed to the
hydrolysis of solid organic substances accumulated at
the lower portion of the reactor.
3.5. Sludge hold-up
Fig. 6 presents the proles of sludge hold-up (in terms
of MLSS and MLVSS), MLVSS per MLSS ratio (VS
ratio) and SVI along the reactor height. At the start of
the experiment, sludge concentration was 40 00045 000
mg MLSS l
1
within the bed portion. After two months
operation at 25C, the sludge concentration (Fig. 6 (D))
at the bed bottom had decreased to 23 000 mg MLSS l
1
and remained unchanged until termination at 13C. At
the middle portion of the bed (20110 cm bed height)
sludge was maintained at a concentration of 30 000
38 000 mg MLSS l
1
for both temperatures. A decrease
of sludge hold-up was observed at the higher portion of
the bed (130150 cm).
SVI, as an index of sludge settlability (Fig. 6 (A)),
exhibited relatively higher values of 40 ml g
1
MLSS at
both the bottom and upper portion of the bed, where
sludge hold-up also exhibited lower values due to the
accumulation of inuent particulate matter. On the
other hand, at the middle portion of the bed, SVI was in
a range of 2030 ml g
1
MLSS owing to the retention of
granular sludge. This observation is consistent with the
tendency of the MLSS and MLVSS proles. Consider-
ing that the SVI of the inoculum granular sludge was 19
ml g
1
MLSS, the deterioration of the settlability of
granular sludge might arise from extended low organic
Fig. 5. Substrate removal pattern along the reactor height. Total VFA
(A); and soluble COD (B). Symbols: (d) 25C; (s) 13C.
Table 2
Eect of temperature on COD reduction
25C 22C 19C 16C 13C
Solid COD removal (%) 83 81 84 77 73
Soluble COD removal (%) 50 51 52 61 45
Total COD removal (%) 70 70 72 71 64
Accumulated COD (g COD m
3
) 68 67 120 78 102
Liqueed fraction
a
(%) 58 56 37 46 33
Methane recovery (%) 60 59 52 49 35
SRT (days) 117 110
a
1 accumulated COD/(inuent solid CODeuent solid COD).
S. Uemura, H. Harada / Bioresource Technology 72 (2000) 275282 279
loading operation. Hence this caused the lowering of
sludge hold-up.
3.6. Sludge characteristics
Fig. 7 presents the proles of C/N ratio, N content,
coenzyme F
420
content, and ATP content of the sludge
along the reactor height at the end of 13C operation
(178th day after start-up). At start-up the value of these
parameters for the seeded sludge were 0.155 mg ATP g
1
MLVSS, 0.132 mg F
420
g
1
MLVSS, 4.61 g C per g N (g
C/g N), and 106 mg N g
1
MLVSS, respectively. Fig.
7(A) obviously shows that the sludge C/N ratio in-
creased along the sludge bed height, whereas the N
content decreased. The F
420
content of the retained
sludge decreased to 1/41/7 as a whole (Fig. 7(B)). Co-
enzyme F
420
is specic for hydrogenotrophic met-
hanogenic bacteria and can be regarded as a parameter
indicating the methanogenic activity derived from hy-
drogen (Dolng and Mulder, 1985). Compared with the
content at the bottom and upper portion of the bed,
hydrogenotrophic methanogenic activity still remained,
to some extent, at the middle sludge layer. The ATP
content (Fig. 7(C)) at a height of 10 cm from the bottom
decreased to about 1/2 of that of the seed sludge, and to
1/4 at a 20130 cm bed height. A value at 10 cm which
was twice as large as that above that height may be
ascribed to the contribution due to microbial cells pro-
vided from the inuent as well as to the eect of growth
of hydrolytic microorganisms capable of decomposing
the solid organics accumulated at the reactor bottom.
The granular sludge as an inoculum was originally
developed from digested sewage sludge harvested from a
municipal sewage treatment plant. The values of pa-
rameters for the digested sewage sludge were 0.041 mg
ATP g
1
MLVSS, 0.015 mg F
420
g
1
MLVSS, 6.4 g C
per g N and 68.7 mg N g
1
MLVSS, respectively. A
similar tendency was observed in the methanogenic ac-
tivity test. The methanogenic activity of the sludge taken
from 10 cm bed height on the 178th day exhibited 0.03
kg CH
4
-COD kg
1
volatile suspended solids (VSS) d
1
with sucrose as a test substrate and 0.08 kg CH
4
-COD
kg
1
VSS d
1
with acetate, respectively. The met-
hanogenic activity for the granular sludge seeded to the
reactor was 0.75 kg CH
4
-COD kg
1
VSS d
1
with su-
crose and 0.77 kg CH
4
-COD kg
1
VSS d
1
with acetate.
On the other hand, values for the digested sewage
sludge, the source of the granular sludge, were 0.07 kg
CH
4
-COD kg
1
VSS d
1
with sucrose and 0.09 kg CH
4
-
COD kg
1
VSS d
1
with acetate. To sum up, the met-
hanogenic activity of the sludge maintained at low
loading (1.6 kg COD m
3
d
1
on average) for six months
was only 4% (with sucrose) and 10% (with acetate) of
that of the granular sludge seeded at the starting point.
Thus, the microbial activity of the granular sludge
seeded to the reactor shifted to the same degree as did
the digested sewage sludge.
Fig. 7. Proles of C/N ratio (A); ATP content (B); F
420
content (C) at
the termination of 13C experiment. Symbols in (A): (d) N content;
(s) C/N ratio.
Fig. 6. Proles of SVI (A); VS ratio (B); MLVSS (C), and MLSS (D).
Symbols: (d) 25C; (s) 13C.
280 S. Uemura, H. Harada / Bioresource Technology 72 (2000) 275282
The SEM observation of the sludge at the end of the
experiment revealed that the sludge remained, as a
whole, still in granular form, but that most of the
granules exhibited pronounced deep cracking, just
looking like an opening mouth. The internal structure of
the seed granules showed a predominance of Met-
hanothrix-like bacteria in lamentous growth. On the
contrary, the interior of the granules fed on sewage ex-
hibited a tendency toward partial autolysis and a low-
ering of cell density due to long-term exposure to
starvation conditions. In particular the long lamentous
Methanothrix-like bacteria broke up into shorter frag-
ments consisting of several cells.
4. Discussion
The feasibility of the UASB process for the treatment
of sewage was investigated at a xed HRT of 4.7 h and
at temperatures ranging from 25C to 13C. The result
showed a satisfactory performance, achieving a total
COD removal of 69.4% based on total inuent and
soluble euent, on average, throughout the entire ex-
periment. As recently reviewed by Seghezzo et al. (1998),
treatment eciencies of sewage by the UASB process
reported so far have varied signicantly, ranging from
7% to 90% of total COD removals within 2202.5 h of
HRT and 732C of temperatures. Our result obtained
in this study shows a relatively good performance
compared to those results. This dierence might be at-
tributed to the following factors: (1) characteristics of SS
(biodegradability, composition, and size distribution),
(2) temperature conditions, (3) SRT applied to the re-
actors, and so forth (Seghezzo et al., 1998).
Fig. 3 and Table 2 show that a considerably large
fraction of CH
4
gas escaped in dissolved form in the
euent. Therefore only the remainder, half of the total
generated CH
4
, could have been recovered. This implied
that the treatment of low strength wastewater such as
sewage results in reducing, to a large extent, the intrinsic
merit of the anaerobic process as an energy recovery
process.
The total COD removal was independent of tem-
perature, but was aected by the inuent solid COD
concentration (Fig. 4). The particulate COD in the in-
uent was virtually removed by means of the entrap-
ment eect of the sludge bed, and the liquefaction rate
of the entrapped organics was prominently dependent
on temperature (Table 2). The fact that the liqueed
fraction of the entrapped solids depends on tempera-
ture might lead directly to an increase of excess sludge
to be treated when the system is operated at lower
temperatures. The entrapment or accumulation of sus-
pended solids within the sludge bed has also been ob-
served by other researchers, not only in UASB reactors
(Barbosa and Sant'Anna Jr., 1989) but also in anaer-
obic uidized bed reactors (Sanz and Fdz-Polanco,
1990). Sanz and Fdz-Polanco (1990) reported a large
accumulation of suspended solids at the top of the
sludge bed in the anaerobic uidized bed reactor
treating municipal sewage. Considering the results of
sludge proles (Fig. 6) together with the C/N ratio
proles (Fig. 7(A)), this suggests that although the inert
and heavier ingredients presented in the inuent were
apt to accumulate near the bottom, solid organic sub-
stances derived from other than biomass increased as
the bed height increased. This can be attributed to the
fact that relatively light particulate organic matter, e.g.
cellulose in the inuent, tended to accumulate at the
upper bed portion.
The microbial activity of the seeded granular sludge
shifted to the same degree as that of the digested sewage
sludge. This could be attributed to the following
factors: (1) the sludge was exposed to starvation con-
ditions, i.e. low loading rate with low strength waste-
water, for six months, and (2) the inert organic
substances derived from inuent sewage accumulated
within the sludge bed, as mentioned above. According
to SEM observation, even though the sludge retained in
the reactor retained its granular form as a whole, the
majority of the granules had a tendency to be partly
broken and collapsed. The Methanothrix-like bacteria
almost disappeared after six-month operation and
shifted from a long lamentous form to short lament
fragments, with a much lower density of viable cells.
This structural change in granules qualitatively sup-
ported the aforementioned phenomena of the shift in
microbial activity.
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