Journal of Biotechnology 45 (I 996) I25- 135

Anaerobic treatment of palm oil mill effluent in a two-stage

up-flow anaerobic sludge blanket (UASB) system
Rafael Borja ap*, Charles J. Banks b, Enrique Sinchez ’
a Institute de lu Grusa (C.S.I.C.). Audu. Padre Gurciu Tejero 4, E-41012 Seuillu, Spin
b Enuironmentul Technology Cenrre, Department ofChemicuf Engineering. Lfniversizy @‘Manchester Institute #‘Science and Technology
(U.M.I.S.T.), P.O. Box 88, Manchester M60 IQD, UK
’ Depurtamento de Estudios de Contaminu&n Ambiental (D.E.CA., C.N.I.C.), P.O. Box 6990, Lo Hubanu. Cuhu

Received 6 June 1995; reviskd 21 July 1995; accepted 11October 1995

Abstract

A high rate anaerobic treatment of palm oil mill effluent (POME) was achieved in a two-stage up-flow anaerobic sludge
blanket (UASB) reactor. The acidogenic reactor acclimated rapidly to the wastewater and was tolerant to a suspended solids
(SS) concentration of 5.4 g 1-r in the influent wastewater. Loading was gradually increased over a period of 100 days
resulting in a satisfactory hydrolysis and acidification giving a maximum rate of acid production of 4.1 g I- ’ d _ ’ acetic acid
at a loading rate of 16.6 g I-’ d-’ COD at a hydraulic retention time of 0.9 days. An increase in alkalinity throughout the
acclimatization maintained the effluent from the reactor at around pH 5.8. The methanogenic reactor was initially fed on
dilutions of the effluent from the first stage reactor after pH adjustment. The loading was gradually increased, and then
stepwise, to 60 g I-’ dd’ COD at which point COD removal efficiency had declined significantly and an accumulation of
long-chain volatile fatty acids was observed. It was concluded that the reactor could work efficiently up to loadings of 30 g
l- ’ dd’ COD, whilst producing a good methane yield and a COD reduction of greater than 90%. Effluent recirculation
alleviated the need for alkali additions to the feed of the methanogenic reactor and a direct coupling of the two reactors was
achieved towards the end of the experimental run of 175 days. Both reactors showed granule formation with distinct
morphological characteristics; these were observed to be formed after 80 days in the acidogenic reactor and after I IO days in
the methanogenic reactor.

Keywordx Palm oil mill effluent (POME); Anaerobic treatment; Two-stage UASB system; pH Control

1. Introduction been given in detail elsewhere (Borja and Banks,

The three main sources of liquid effluent gener-
ated from a conventional palm oil mill are sterilizer
condensate, hydrocyclone waste and separator sludge.
The process flow-sheet for palm oil extraction has
* Corresponding author.
1994).
For a well-controlled conventional mill, about 0.9,
0.1 and 1.5 m3 of steriliser condensate, separator
sludge and hydrocyclone waste are generated for
each tonne of crude palm oil produced. In most
mills, these three wastewater streams are combined
together resulting in a viscous brown liquid contain-
ing fine suspended solids, the characteristics of which

0168- 16.56/96/$15.00 0 1996 Elsevier Science B.V. All rights reserved

SSDI 0168-1656(95)00154-9
126 R. Borju et aI./ Journal of’Biotechnology 45 (1996) 125-135

Table I retention time but ran into operational problems of

Properties of palm oil mill effluent (POME) poor sludge settleability. Chin (1981) described a
Parameter a Sterilizer Hydrocylone Centrifuge Mixed series of bench-scaie experiments comparing full
effluent effluent effluent effluent
mixed continuous stirred tank reactor (CSTR) di-
BOD IO-25 - 17-35 I l-30 gesters with similar digesters operating with a cell
COD 30-60 - 40-75 30-70
recycle system; for a 90% COD removal a hydraulic
Total solids 40-50 5-15 35-70 30-65
5-12 12-18 9-25
retention time (HRT) of 20 d was required. Ibrahim
Suspended 3-5
solids et al. (1984) showed that at 45°C an anaerobic
Oil 2-3 l-5 5-15 5-13 contact digester could be operated at a maximum
Ammoniacal 0.02-0.08 - 0.02-0.08 0.02-0.08 organic load of 6.2 g 1-l d-’ COD, giving a per-
N centage reduction of around 94%; a higher loading
Total N 0.35-0.60 0.07-0.15 OS-O.9 0.5-0.9 of 7.0 g 1-l dd ’ could be treated but only at the
PH 4.5-5.5 - 3.5-4.5 3.5-4.5
higher temperature of 50°C. Cail and Bat-ford (1985)
a In g I-‘. used a semicontinuous mesophilic anaerobic digester
with a volumetric loading of 12.6 g I-’ d- ’ (HRT of
are shown in Table 1. If discharged untreated this 6 d). Ng et al. (1985) showed 85% reduction in
effluent can cause considerable environmental prob- pollution load using a two-stage anaerobic digestion
lems (Davis and Reilly, 1980). system with no cell recycle and a total HRT of 31 d.
The increasingly stringent water quality regula- The data of Edewor (1986) showed that a IO-day
tions being introduced in many countries have forced retention time in a single-stage anaerobic contact
factories to investigate a wide range of approaches digester gave a COD removal efficiency of 93.8%. A
for the treatment of palm oil mill effluent (POME) two-stage variant of this system gave an average
and related wastewaters (Petitpierre, 1982). These removal efficiency of about 96% for HRTs between
include: simple skimming devices (Roge and Ve- 7 and 15 d. The use of an up-flow anaerobic sludge
layuthan, 1981; Ng et al., 1988); land disposal (Ma blanket (UASB) reactor for treating this wastewater
and Ong, 1986); use as animal fodder (Sutanto, has been reported in a previous paper (Borja and
1983; Ma and Ong, 1986); ultrafiltration (Suwandi, Banks, 1994). Over 80% COD was removed at
1981; Turisin et al., 1981); chemical coagulation and loading rates up to 10.6 g 1-l d-’ COD, and at the
flotation (Badri, 1984; Ho and Chan, 1986; Chin et highest influent concentration (42.5 g 1-l > reactor
al., 1987; Ng et al., 1987a; Karim and Hie, 1987); instability was observed.
and various aerobic (Abdul et al., 1989) and anaero- Although the UASB reactor has been successfully
bic microbiological processes. Anaerobic biological applied to a wide range of both low and medium
systems offer greater potential for the treatment of strength food industry wastewaters the mechanism of
POME, as they do not have the high energy demand bacterial granulation is still not fully understood
of aeration as required by aerobic biological systems (Lettinga et al., 1980; Lettinga and Vinken, 1980;
(Edewor, 1986; Ng et al., 1987b; Ma and Ong, Goodwin et al., 1992). The process appears to be
1988). The use of conventional anaerobic lagoons particularly sensitive to the suspended solids (SS)
(Hum et al., 1981; John, 198.5) or digesters (Ho and loading which can be applied, and also to the nature
Tan, 1985) to treat POME is characterized by long of the organic content of the wastewater. Souza
residence times, often in excess of 20 d, necessitat- (1986) reported that for a single-stage UASB reactor
ing large areas of land or large digesters. The appli- then a value of 0.04 g SS g- ’ COD could be treated
cation of modem high rate anaerobic digester tech- and provided the ratio was not exceeded then sus-
nologies (Pandey and Ramakrishna, 19901, such as pended solids concentrations as high as 1 g 1-I could
up-flow or down-flow filters, fluidized beds, up-flow be applied.
anaerobic sludge blanket (UASB) systems or up-flow At high organic loadings or at low temperatures
floe digesters, for the disposal of POME is rare. the insoluble organic fraction of the wastewater tends
Peyton et al. (1979) used a laboratory scale to accumulate within the granule or sludge blanket
mesophilic contact stabilization process with a IO-day region of the reactor; this leads to granule destabi-
 R. Borju et ul./JournuI
lization or inhibition of granule formation (Hulshoff
Pol and Lettinga, 1986; Vigneswaran et al., 1986).
Under these conditions methanogenesis is affected,
leading ultimately to reactor failure as a result of
increased acid concentrations. One solution is to
remove the solids from the wastewater stream prior
to it entering the reactor as was the case in previous
studies using POME. This is now, however, ideal for
two reasons. Firstly, the suspended organic solids
have a high potential for gas production making
process economics more favourable. Secondly, re-
moval prior to UASB treatment generates a sludge
which requires disposal at additional cost.
Hydrolysis and acidification of organic solids in a
separate stage to that of methanogenesis was first
proposed by Ghosh et al. (1975) and two-stage reac-
tor systems, using conventional continuous stirred
tank reactors (CSTRs), are not uncommon. To apply
the principal of a two-stage treatment using a pair of
upflow anaerobic reactors is the subject of this re-
search, the objective being to evaluate the effect of a
staged treatment on sludge granulation.
2. Materials and methods
2.1. Experimental design
The schematic diagram of the two-stage UASB
system used in this study is shown in Fig. 1. A 12-l
A: Acidogenic UASB reactor.
M: Methanogenic UASB reactor.
R: Refrigerated feed tank.
Fig. I. Schematic diagram of the two-stage UASB system used in this study.
of Biorechnology 45 (1996) 125-135 127
UASB reactor (A) with 13 cm i.d. and a 5-1 UASB
reactor (MI with 9 cm i.d. were used for the acido-
genie and methanogenic reactions of a two-stage
process. The reactors were separated by a 2.5-l bal-
ancing tank receiving the effluent from reactor A and
providing the feed reservoir for reactor M. The
reactors and balancing tank were maintained at a
constant temperature of 35°C in an insulated cabinet.
Each reactor was of a typical UASB design having a
gas-biomass-liquid separator at the head of the
column and an influent liquid distributor at the base.
Each column was connected to a water displacement
type gasometer filled with an acidified brine solution
to prevent CO, dissolution. The reactor contents
could be sampled at various heights along the col-
umn by means of a series of six sampling ports along
its length. A detailed design description of this type
of small-scale UASB reactor has been given else-
where (Borja and Banks, 1994).
2.2. Wastewater
The reactors were fed on dilutions of a mixed
waste stream (POME) typical of that produced in the
palm oil manufacturing process. The features and
composition of the POME are summarized in Table
2, the data shows average values from five separate
determinations for each parameter; the deviation be-
tween these repetitive analyses was less than 3%.
During the initial feeding of the wastewater to the
EPFLUENT
RECIRCULATION
S: Separation tank.
P: Periststtic pump.
G: Gas collector.
128 R. Borju er ai./Journal oj’Biorechn&gy 45 (19961 125-135

Table 2 the first 100 days of operation an influent SS concen-

Composition and features of the POME used tration of 5.4 g 1-l was maintained by diluting
PH 4.4 POME with tap water. Between days 101 to 120 the
COD(g I-‘) 30.6
influent SS concentration was increased to 10.8 g 1-l
Suspended solids (SS; g l- ’ ) 10.8
8. I by feeding with undiluted POME. From the 121st
Volatile suspended solids (VSS; g 1.. ’ )
Total solids (TS; g I- ’ ) 31.2 day the concentration was dropped once again to 5.4
Volatile solids (VS; g I- ’ ) 24.3 g 1-l SS. The initial loading rate to reactor M was
Kjeldahl N (mg I _ ’ ) 365 1.1 g I-’ d- ’ COD and this was achieved by
P(mg 1-l) 110
dilution with tap water into the balancing tank so as
Fe(mg 1-l) 205
60
to prevent inhibition by volatile fatty acids. As re-
S(mgl-‘1
Ca(mg I-‘) 220 moval efficiency increased above 80% mark the
Na(mg I-‘) 4 loading was gradually increased over a 60-d period
K(mgl-‘) 510 so as to maintain this level of treatment. At the end
Mg(mg 1-l) 170
of this time tap water dilution was no longer neces-
Cu(mgl-‘) I
6
sary but pH adjustment continued until day 140. At
Zn(mgl-‘)
Mo(mp I-‘) 0. I this time alkali additions to the balancing tank were
Co(mg 1-l) 0.007 stopped and after a further 5 d (day 1461, the balanc-
Mn(mgl-‘) 0.6 ing tank was removed and reactor A effluent was fed
Ni(mgl-‘) I .2
directly to reactor M.
Al(mgl-‘1 120
B(mgI-‘) 0.9
In an attempt to prevent a loss in alkalinity from
Ba(mg I-‘) 0.3 the system after alkali additions had ceased, the
Si (mg l- ’ ) 55 effluent from M was recirculated into reactor A from
day 160 onwards.

reactors the pH of the POME was adjusted to 6.0
using a 5 N sodium hydroxide solution. Different
dilutions of the POME were prepared using tap
water so as to cover a range of suspended solids
concentrations.
2.3. Seed sludge
Reactor A was inoculated with sludge taken from
a completely mixed acidogenic reactor which was
part of a two-stage system treating concentrated ice-
cream wastewater. Reactor M was seeded with
methanogenic sludge taken from a CSTR reactor
treating POME. The initial sludge concentrations
were 18.8 and 16.1 g 1-l of volatile solids (V’S) for
reactors A and M, respectively.
2.4. Loading regime
Reactor A was started with an initial loading rate
of 2.3 g 1-l dd ’ COD and a HRT of 6.5 d. There-
after, the loading rate was increased stepwise up to
17.3 g I- ’ d- ’ COD over a period of 120 d. During
2.5. Sampling and chemical analyses
Gas volume and composition were monitored daily
and effluent samples from both reactors were anal-
ysed for COD, pH, and VFA (volatile fatty acids)
every 3-4 days; the analytical methods used were in
accordance with standard methods (American Public
Health Association, 1985).
Methane was determined by gas chromatography
using a stainless steel column (200 X 0.3 cm) packed
with active carbon (30-60 mesh), the gas flow pass-
ing over a thermal conductivity detector.
Volatile fatty acids (VFA) were determined using
a gas chromatograph equipped with a 2 m X 4 mm
glass column packed with Supelcopor (loo- 120
mesh) coated with 10% Fluorad FC 431. The tem-
peratures of the column, the injection port and the
flame ionization detector were 130, 220 and 240°C.
respectively. Nitrogen saturated with formic acid was
used as the carrier gas at a flow rate of 50 ml min - ’ .
The POME was digested in aqua regia, and its
elemental composition analysed by inductively cou-
pled plasma (ICP) emission spectrophotometry.
 R. Borju et d/Journal
The structure of the granular sludge was made
with scanning electron microscope @EM).
3. Results and discussion
3.1. Stage separation
Biogas composition and volatile acid concentra-
tion indicated that the stage separation was easily
achieved in an early stage. Adjusting pH seems
simpler than other reported techniques of stage sepa-
ration, such as membrane dialysis, selective inhibi-
tion and kinetic control (Massey and Pohland, 1978).
In addition, a pH of around 6.0 appears to be the
optimum for acidogenic activity. If the pH is not
controlled the acidogenic biomass tends to buffer
itself to a pH that depends on the other environmen-
tal conditions in the reactor, but mainly on the nature
of the wastewater (Zoetemeyer et al., 1982; Hanaki
et al., 1987).
3.2. Pelfonance of the acidogenic reactor
From the mass balance of volatile fatty acids in
reactor A the rate of acid production, in terms of
-1
HAc formation rate (g I d )
i
5 15
COD loadinrrate
L---ff Acid formation
Fig. 2. The response of the acidogenic reactor, in terms of the gas (1 d- ’ ) and acid (g I- ’ d- ’ HAc) production rates, to organic loading (g
I-’ d- ’ COD). Th e points at the highest loading value show a depression in rates due to the high influent suspended solids concentration
(10.8 g I-‘).
of Biotechnology 45 (1996) 125-135 129
acetic acid equivalents, can be determined and ex-
pressed as g acetic acid per 1 reactor volume and day.
As can be seen from Fig. 2, both the rate of acid and
gas production showed a similar trend with increases
in both parameters as organic loading increased in
the range 2.5-16.6 g l- ’ d- ’ COD. This corre-
sponds to the time during which the reactor was
operated with an influent SS concentration of 5.4 g
I-‘; the maximum acid production was 4.1 g 1-l
d- ’ acetic acid and corresponded to a HRT of 0.9 d.
When the influent SS concentration was increased
to 10.8 g 1-l between days 101-120 both the acid
and gas production rates decreased sharply; this cor-
responds to an organic loading rate of 17.3 g I-’ dd ’
COD. During this period of operation the height of
the sludge blanket increased and can be attributed to
an accumulation of non degraded organic solids
within the reactor. This in turn led to a decreased in
acidogenic activity which may have been as a result
of the poor mixing observed within the sludge blan-
ket at this time. The gas produced in the acidogenic
reactor was primarily carbon dioxide although at the
lower organic loadings methane was detected at lev-
els up to 9%. As the organic loading increased
-1
Gas production (I d-j
- 18
1
16
14
12
10
0
20
(g ;;‘I
rate *- Gas production
130 R. Borju et al./ Journal ofBiotechnology 45 (1996) 125-135

methane concentration decreased and hydrogen be-
came detectable and was consistently measured be-
tween 1 and 2% from day 80 onwards. The results of
the gas analysis are in line with expectations and the
appearance of hydrogen is indicative of the subse-
quent breakdown of longer chain volatile fatty acids
to acetic acid and the loss of autotrophic
methanogenic activity. During the first 100 days of
operation the reactor alkalinity gradually increased
from an initial value of 2100 mg l- ’ to a final value
of 5200 mg 1- ’ as CaCO, at loading rate of 16.6 g
1-l d-’ COD. The pH at the top of the sludge
blanket remained relatively constant, between 5.2 to
5.8 over this time period showing that the increased
acid production was being satisfactorily compensated
by an increased in alkalinity.
Between days 120- 175 the acidogenic reactor
was fully acclimated and operating with a influent
SS concentration of 3.8 g l-‘, it was however recov-
ering from the high loading and solids concentration
applied between day 1OO- 120. The predominant VFA
found during this period was acetic acid (Fig. 3) and
accounted for between 45 to 60% of the total acid
during this period. The level of total mixed acids
dropped over a 45-d period from 5.6 g 1-l to just
over 4.0 g l-‘, and the proportion of acetic acid to
that of longer chain acids increased slightly during
this period showing recovery to conditions similar to
those observed shortly before day 100. Propionic,
butyric and valeric acids were all detectable in con-
centrations greater than 100 ppm. Free VFAs as
products of the acidogenesis, can diffuse into the
cells of anaerobic bacteria and ionise to decrease the
pH. This is generally recognized as inhibiting acido-
genesis (Dinopoulou and Sterrit, 1988). The alkalin-
ity of the influent allows the neutralization of the
free VFAs, preventing the fall in pH from its opti-
mum value both in the reactant influent and in the
cells, while the VFAs in the acidogenic UASB in-
creased.
3.3. Performance of the methanogenic reactor
The performance of the methanogenic reactor was
monitored using the COD removal efficiency (%I,
the specific gas production rate (1 CH, gg ’ COD
removed), and the methane content of the biogas
(%I; all of these are known to respond to conditions
of organic overloading and are indicative of the
system stability.

VFA concentration (g i’)

61 ~~
1

I I
1” ---- +&_“-_._--_
i’ I
0 --I-27? L i
100 110 120 130 140 150 160 170 180 190 200
Operation time (d)

- c2 c3 - c4 " c5 . Total VFA (

~_
Fig. 3. The volatile fatty acid profile in the acidogemc reactor following the period of maximum loading (days lOI- 119).Total acids are
expressed as acetic acid equivalents (g I- ’ HAc) and individual acids in g I- ’
 R. Borja et al./ Journal of Biotechnology 45 (19%) 125-135 131

-1 -1
A (%I; 6 (%I C, biogas (I I d 1; D, COD (g l-b-j
lOOr-
F70

60

1
50

40

30

+$y::2$Q
+-e -9
-~~~ i_ ~__ -L- ___-
20

10

0
50 100 150 200
Operation time (d)
r--
-. A --t B + C --ff D

Fig. 4. Response of the reactor performance indicators in the methanogenic reactor to a gradual, and then stepwise, increase in organic load
over 150 d. The last 25-d period of operation is without alkali additions to the influent and with and effluent recycle to the acidogenic
reactor. (A) Methane content of the biogas (%); (B) COD removal efficiency (%); (C) specific biogas production rate (I I- ’ d- ‘); (D)
organic loading rate (g I- ’ d- ’ COD). -

VFA concentration (g (‘1

4r---------
1

0 i ._~___1__ _._ L

100 110 120 130 140 150 160 170 180 190
Operation time (d)
I

-- c.2 +-C3 --t- c4 ifc5 - Total WA

Fig. 5. Total volatile fatty acid (g I- ’ HAc)and volatile fatty acid components (g I - ’ ) in the methanogenic reactor during the period from
days 120 to 175.
132 R. Borju et al./ Journal c$Biotrchnolo~y 45 (1996) 125-13.5

With any methanogenic reactor it is necessary to
assay the biodegradative capacity with respect to
increasing organic load so as to optimise reactor size
whilst maintaining treatment efficiency. To achieve
this the organic load must be either gradually in-
creased or increased stepwise over a period of time
to allow acclimatization of the microbial population
and pseudo steady state conditions to be reached at
each loading value. The reactor showed a rapid
initial adjustment to the POME feed reaching 90%
COD removal efficiency at the initial loading of 1.1
g I- i d- ’ within 10 d (Fig. 4). The increases in load
applied were of gradual nature over the first 120
days of operation cumulating in a loading rate of
22.9 g I- ’ d- ’ COD. During this period of operation
biogas production increased in line with increases in
loading, whilst the percentage methane content of the
biogas gradually decreased from an initial value of
over 80% to one of around 65%. Although there was
some variation in COD removal efficiency this aver-
aged over 90% with low values on occasions of less
than 80%, between days 121 and 149 two sharp
increases in loading were applied, the first taking the
load to 45 g 1-l d- ’ COD and the second to 60 g
1-l dd ’ COD. During this period the gas production
rate also rose to a maximum value of 26 1 1-l d- ‘,
whilst the COD removal efficiency showed a steady
decline to below the 80% level.
At the same time the volatile fatty acid level in
the effluent from the reactor rose to 3 and then 3.5 g
1-l mixed acid for the loadings of 45 and 60 g I-’
d- ’ COD, respectively (Fig. 5). Of the mixed acids
present propionic was dominant with levels in excess
of2gl_1, indicating a build up of longer chain acids
and rate limitation in the acidogenic conversion stage.
Such conditions normally lead to a rapid souring of
the anaerobic process with eventual inhibition of
methanogenic activity. For this reason the loading
rate was again reduced to 28.2 g 1-l d- ’ COD for
the next 20 d before dropping to a loading of 20.0 g
I-’ d-’ for the final days of the experimental run.
COD removal efficiency improved reaching over
90%, whilst the gas production rate dropped back to
10 I I- ’ d- ‘. The volatile fatty acid levels in the
reactor effluent rapidly fell to a level of around 0.5 g
I-‘, with the C3, C4, and C5 chain length acids
being removed to concentrations of less than 0.1 g
I-‘; this was indicative of a healthy sludge and
restoration of acetogenic activity. Overall, the
methane yield throughout the course of the experi-

Alkalinity (mg i’)

8000 , ..

6000 i

5000

I--- ! .
4000
140 145 150 155 160 165 170 175 180
Operation time (d)
Fig. 6. Change in alkalinity (mg I ‘, as CaCO,) in the methanogenic reactor on cessation of alkali addition to the influent. An effluent
recycle to the acidogenic reactor was initiated on day 155at a ratio of 0.33 parts reactor M effluent to I part reactor A feed.
 R. Borja et cl!./ Journal of Biotechnology
ment ranged from 0.30-0.33 1 CH, per g COD,,,,,,,,
this is a little less than the theoretical value of 0.35.
On day 140 alkali addition to reactor M feed was
ceased and 5 days later the balancing tank between
the reactors was removed. As can be seen from Fig.
6 the alkalinity of the system steadily dropped from
7.0 g 1-l to less than 6.0 g 1-l over the following 10
d. In order to maintain alkalinity within the system a
recycle of effluent from reactor M to reactor A was
installed with a recirculation ratio of 0.33. This was
successful in preventing any further loss of alkalinity
which stabilized at around 5.3 g 1-l giving an
influent feed pH of 6.0 to reactor M. At an organic
loading less than 28.2 g 1-l d- ’ COD the ratio of
volatile acid to alkalinity could be maintained in the
range 0.1-0.2 which is suitable for stable digestion
and methanogenesis.
3.4. Sludge granulation
Both reactors were seeded with non granular
sludge from single-stage digesters and as such were
acclimated gradually at low loading rates to the new
feedstock and reactor conditions. The first signs of
granulation of the sludge were in the acidogenic
reactor on day 80 when the influent SS concentration
and the loading rate were 5.4 g 1-l and 16.6 g 1-l
dd ’ COD, respectively. The granules were 2-3 mm
in size and had an offensive odour. Granulation in
the methanogenic reactor was found after 110 days
of operation, when the loading rate was 22.1 g 1-l
d- ’ COD, and the granules had an average diameter
of l-2 mm.
The granules that formed in the methanogenic and
acetogenic reactors differed in a number of ways
other than those mentioned above. The acidogenic
granules were much more fragile, appeared less
dense, and had a much lower settling velocity. Mor-
phological differences were apparent on examination
using SEM. The methanogenic granules were similar
to those previously reported in UASB reactors treat-
ing food industry wastewaters (Wiegant and Let-
tinga, 1985; Dolfing et al., 1986; Dinopoulou and
Sterrit, 1988; Goodwin et al., 1992) in that they
consisted of networks of long multicellular filaments,
typical of Merhanothrix spp., with a diverse selec-
tion of rod and coccus type bacteria entrapped in a
dense matrix. The process of granulation under aci-
45 (1996) 125-135 133
dogenic conditions is far less studied than in mixed
digestion although it has been reported by a number
of authors (Cohen and Zoetemeyer, 1979; Lettinga et
al., 1980). The acidogenic granules found in this
study were devoid of the multicellular filaments and
appeared to be bound together by chains of a large
rod-shaped bacterium with the surface of the granule
predominantly consisting of short plump rods and
various coccus types. There appeared to be a smaller
variety of morphological forms present within the
acidogenic granules than in the methanogenic ones,
and the density of packing of the bacteria within the
methanogenic granules appeared a lot higher; the
latter is likely to effect strength and settlement char-
acteristics (Prensier et al., 1988).
Sludge granulation is a dynamic process without a
clear starting point and the factors which initiate this
process and influence granular formation are not
fully understood (Dolfing et al., 1986; Colleran,
1988). One factor which is thought to be important is
the superficial biogas developing rate and these were
calculated at the time the granules were formed in
both of the reactors. The values found were 1.7 and
5.4 m dd ’ for the acidogenic and metbanogenic
reactors, respectively, and as such the value for the
acidogenic reactor is less than previously reported
rates which lie in the range 3.6 to 10 (Wu et al.,
1985; Wiegant and Lettinga, 1985). This result was
not unexpected as overall the rate of gas production
in an acidogenic reactor is much lower than in a
methanogenic reactor as the reaction products (VFA)
are liquid with only small quantities of H, and CO,
being evolved.
4. Conclusions
The results clearly show that it is possible to split
the anaerobic digestion process into two distinct
stages in a pair of upflow anaerobic sludge blanket
reactors. The acidogenic reactor, although tolerant to
an influent suspended solids concentration of 5.4 g
1-l) showed signs of reduced solids liquification
when the loading was increased and the suspended
solids concentration was 10.8 g l- ’ .
Based on the efficiency indicators used it is clear
that the methanogenic reactor can quickly acclimate
to a feed which comprises the effluent from the
134 R. Borju et al. / Journd
acidogenic reactor. Although loadings as high as 60
g 1-l dd’ COD were applied these resulted in a
significant loss in COD removal efficiency and car-
bon flow from longer chain fatty acids to methane. A
maximum load of around 30 g 1-l d-’ COD should
ensure an overall 90% reduction in COD and effi-
cient acid conversion to methane.
Recirculation of effluent from the methanogenic
reactor to the acidogenic reactor proved to be an
effective way of maintaining reactor pH around 6
which was suitable for efficient methanogenesis; this
would save on the addition of chemicals and reduce
overall running costs on a large-scale process and
also alleviate the need for an intermediate balancing
tank between the reactors.
Granulation was observed in both reactors but
their morphological characteristics were distinct. The
mechanism of granulation was not elucidated but
was shown to occur at typical values of superficial
biogas developing rates in the methanogenic reactor
and at values lower than previously reported in the
acidogenic reactor.
Acknowledgements
The authors wish to thank the ‘Departamento de
Postgrado y Especializacion de1 CSIC’ (Spain) for
financial support granted for the realization of this
work.
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