Bioresource Technology 71 (2000) 143±149

Anaerobic pre-treatment of slaughterhouse wastewater using

®xed-®lm reactors
R. del Pozo, V. Diez*, S. Beltr
an
Department of Biotechnology and Food Science, Faculty of Food Science and Technology and Chemistry Science, University of Burgos,
Pza. Misael Ba~ nuelos s/n, 09001 Burgos, Spain
Received 30 December 1998; received in revised form 14 April 1999; accepted 21 April 1999

Abstract
The aim of this work was to study the performance of anaerobic ®xed-®lm reactors with non-random support, for poultry
slaughterhouse wastewater pre-treatment, including the in¯uence of operating conditions. The work was carried out with two lab-
scale reactors, one up¯ow and the other down¯ow, both equipped with vertical corrugated PVC tubes as support and a recirculation
circuit. Both reactors were operated at 35°C.
COD removal eciencies ranging from 85% to 95% were achieved for organic loading rates of 8 kg COD mÿ3 dÿ1 , while the
highest organic loading rates (35 kg COD mÿ3 dÿ1 ) led to eciencies of 55±75%. The reactors did not show destabilization after 12 h
shock loads of 50 kg COD mÿ3 dÿ1 .
Reactor stability was easily achieved under intermittent operation, with weekend breaks, after which the reactors rapidly re-
turned to their optimal performance. The in¯uences of the hydraulic retention time, temperature, the recirculation ratio and ¯ow
direction were also studied. Ó 1999 Elsevier Science Ltd. All rights reserved.

Keywords: Anaerobic pre-treatment; Slaughterhouse wastewater; Fixed-®lm reactors

1. Introduction treatment obtaining Chemical Oxygen Demand (COD)

The slaughterhouse industry is a very old human
activity and, although it is still a relatively small-scale
industrial sector, its environmental impact has grown
considerably due to the increase in plant production.
The main pollutant in slaughterhouse e‚uents is bio-
degradable organic matter in such a high concentration
that aerobic treatment processes are limited by the high
energy consumption needed for aeration, the oxygen
transfer capacity and the high sludge production. This
type of wastewater also has a high grease concentration,
which makes suspended bio-mass anaerobic reactors,
such as the Up¯ow Anaerobic Sludge Bed (UASB), also
unsuitable. This is due to lipid accumulation over the
¯ocs (Chen and Shyu, 1998) with the consequent loss of
eciency, and because of desgasi®cation limitations
which can cause ¯otation and biomass wash-out (Sayed
et al., 1987).
Anaerobic Filters (AF) with random support have
been successfully used for slaughterhouse wastewater
*
Corresponding author. Tel.: +34-947-258810; fax: +34-947-258831;
e-mail: vdiezb@ubu.es
removal eciencies between 80% and 90% for organic
loads up to 20±25 kg COD mÿ3 dÿ1 , and a quick start-
up of two or three weeks (Henze and Hamerroes,
1983; Tritt, 1992; Borja et al., 1993). In addition, ad-
equate performance under acidifying operating condi-
tions has already been reported for this type of reactor
(Borja and Banks, 1994) which have also shown sat-
isfactory stability when treating highly loaded waste-
water under sudden changes in the operating conditions
such as temperature shocks, organic and hydraulic
overloads and feed interruptions (Metzner and Temper,
1990). In spite of this, random-support ®lters require
periodical wash-outs to avoid reactor clogging. For
this reason di€erent support options should be con-
sidered.
The aim of this work was to evaluate the performance
of Anaerobic Fixed-Film Reactors (AFFR) with non-
random support in the digestion of slaughterhouse
wastewater. The experiments carried out were designed
to study the in¯uence of the organic loading rate (Bv ),
the hydraulic retention time (HRT), the in¯uent chem-
ical oxygen demand (COD), temperature, recirculation,
¯ow direction and intermittent operation.

0960-8524/00/$ ± see front matter Ó 1999 Elsevier Science Ltd. All rights reserved.
PII: S 0 9 6 0 - 8 5 2 4 ( 9 9 ) 0 0 0 6 5 - 6
144 R. del Pozo et al. / Bioresource Technology 71 (2000) 143±149
2. Methods
2.1. Anaerobic ®xed ®lm reactors
Two lab-scale tubular-shaped reactors, built in
Plexiglas, 80 cm high and 8 cm in diameter were used in
this work. PVC corrugated tubes; 70 cm high and 2.5 cm
in diameter were employed as non-random supporting
material. They provided a super®cial area for biomass
attachment of approximately 250 m2 mÿ3 and were ar-
ranged vertically in order to avoid reactor plugging due
to the high solids concentration of the slaughterhouse
wastewater (950 mg lÿ1 ). The only di€erence between
both reactors was the direction of ¯ow. Water circulated
upwards in the U-AFFR and downwards in the D-
AFFR, as shown in Fig. 1.
Both reactors had a recirculation circuit in order to
control super®cial water velocity and mixing conditions.
Temperature was maintained at 35°C with a thermo-
static bath. Biogas production was measured with an
electromechanical gas-meter device described in detail
elsewhere (Diez, 1991). Periodical re-calibration was
Fig. 1. Flowchart of the anaerobic ®xed-®lm reactors: (1) up¯ow-AFFR; (2) down¯ow-AFFR; (3) feed; (4) e‚uent; (5) recirculation; (6) gas-meter
device (L.C.: level control, C.: counter); (7) thermostatic jacket.
needed due to electrode corrosion caused by the H2 S
content of the biogas.
2.2. Slaughterhouse wastewater
The wastewater used in this work was taken from the
poultry slaughterhouse of Cooperativa Avõcola y
Ganadera de Burgos, a local poultry processing plant.
Chicken blood was used to increase feed COD concen-
tration when needed. The feed tank was stirred with an
internal recirculation system and the input to the reactor
was placed so as to avoid collection of the settled solids
and to provide a homogeneous suspended solids con-
centration in the in¯uent. Table 1 presents the usual
characteristics of poultry slaughterhouse wastewater.
2.3. Reactor operation
Both reactors were operated for 13 months, stopped
for 4 months and then operated again for 4 more
months. Despite such a long stop, the reactors were
quickly and satisfactorily re-started. After the in¯uence
 R. del Pozo et al. / Bioresource Technology 71 (2000) 143±149 145

Table 1 and 12 kg COD mÿ3 dÿ1 . The COD removal achieved

Poultry slaughterhouse wastewater composition
Interval
Q (m3 tonÿ1 )a 3.4±15
BOD5 (mg lÿ1 ) 600±1700
COD (mg lÿ1 ) 1100±3400
TSS (mg lÿ1 ) 600±1900
TKN (mg lÿ1 ) 100±300
TP (mg lÿ1 ) ± 70
Grease (mg lÿ1 ) ± 110
a
Ton of broiler.
of ¯ow direction was studied, the D-AFFR was operated
with a week-end break program, a very common oper-
ating procedure in small slaughterhouses, while the U-
AFFR was fed continuously. Table 2 shows the organic
loading rate (Bv ) and the in¯uent organic matter con-
centration (CODf ) as well as the reactor employed for
the study of each operation parameter.
2.4. Analytical procedures
Feed and e‚uent samples were analyzed daily for
chemical oxygen demand (COD) and pH. Alkalinity
ratio (IA:PA), de®ned as the intermediate alkalinity (IA;
titration from pH 5.75 to 4.3) divided by partial alka-
linity (PA; from original pH to 5.75) (Ripley et al.,
1986), was also measured daily. Total and volatile sus-
pended solids (TSS, VSS), total kjeldhal nitrogen (TKN)
and ammoniacal nitrogen (NHx ±N) concentrations were
determined weekly, increasing the frequency when re-
quired. All analyses were developed following Standard
Method's procedures (APHA, 1998). Finally, two bio-
gas production measurements were taken daily.
3. Results and discussion
3.1. In¯uence of the direction of ¯ow
Both reactors (U-AFFR and D-AFFR) were oper-
ated for 130 days at a hydraulic retention time (HRT) of
6 h and moderate organic loading rates (Bv ) between 2
for the two reactors employed is presented in Fig. 2 as a
Average function of the organic loading rates. A statistical t-test
8 with a con®dence level (a) of 0.05, showed that there was
1200 no di€erence in COD removal eciency between both
2100 reactors.
950 Total suspended solids (TSS) values for the two re-
220
actors ¯uctuated within a wide interval, between 60 and
220 mg lÿ1 , although both reactors had about the same
average TSS concentration; 137 for the U-AFFR and
146 for the D-AFFR. No relationship was found be-
tween this ¯uctuation and any other operating condi-
tion, therefore, it was attributed to the design of the
output pipes in such small reactors as those employed in
this work.
Accumulation of suspended biomass was observed in
both reactors during the complete period of study pre-
sented in Table 2. After 17 months of operation, volatile
suspended solids (VSS) concentrations were 6800 mg lÿ1
for the U-AFFR and 5000 mg lÿ1 for the D-AFFR. In
both cases, suspended biomass formed easily settling
¯ocks.
3.2. In¯uence of the organic loading rate (Bv )
3.2.1. In¯uence of a progressive load increase
In order to study the in¯uence of the organic loading
rate on process eciency, values between 5 and 35 kg
COD mÿ3 dÿ1 were applied. Fig. 3 shows that a decrease
of COD removal eciency, calculated with e‚uent
centrifuged COD, occurred when the organic loading
rate in the up¯ow reactor increased, in other words,
when Bv rose from 8 to 30 kg COD mÿ3 dÿ1 , COD re-
moval decreased from 85±95% to 55±75%. An increase
in the organic loading rate slightly a€ected the
ammoni®cation rate of the anaerobic reactor, lowering
the NHx ±N/TKN ratio of the e‚uent from 96% to 85%.
Organic loads used in this work were considerably
higher than those found in the literature for this type of
reactor, the highest of which was 25 kg COD mÿ3 dÿ1
(Borja et al., 1995). Under such conditions and despite
the decrease in removal eciency, the pH was above 7.4
and the alkalinity ratio (IA:PA) below 0.54, which in-

Table 2
Operating conditions
Operation parameter studied Reactor Bv (kg COD mÿ3 dÿ1 ) CODf (mg lÿ1 )
Direction of ¯ow U, D-AFFR 7.7 1800
Organic loading rate U-AFFR 5±35 4000 and 11800
Shock organic load U, D-AFFR 50 6000
Feed COD concentration U-AFFR 9 2560±11300
Hydraulic retention time D-AFFRa 27 2500
Intermittent operation D-AFFRa 7.7 1600
Recirculation D-AFFRa 9.6 2060
Temperature D-AFFRa 9.4 2160
a
Intermittent operation with a break program of 60 h per week.
146 R. del Pozo et al. / Bioresource Technology 71 (2000) 143±149

Fig. 2. COD removal in the up¯ow and down¯ow AFFR with moderate organic loads (calculated with e‚uent total COD concentration).

Fig. 3. COD removal in the up¯ow-AFFR versus organic loading rate (Bv ) (calculated with e‚uent centrifuged COD concentration).

dicated a tolerable accumulation of Volatile Fatty Acids
(VFA). According to Ripley et al. (1986), although op-
timal anaerobic digestion occurs at IA:PA values from
0.25 to 0.3, methanogenic inhibition does not start until
0.8.
3.2.2. E€ect of transient organic overloads
Shock loads are common in all kinds of industries
and they may have an important harmful e€ect on an-
aerobic biological processes, causing destabilization of
the microbial populations. This leads to VFA accumu-
lation that can acidify the reactor and therefore inhibit
methanogenic microorganisms. In this case the reactor
enters an accumulation±inhibition cycle which implies
its total collapse.
A shock load of 50 kg COD mÿ3 dÿ1 maintained for
12 h, with a hydraulic retention time around 3 h was
applied to the U-AFFR. Fig. 4 shows the evolution of
 R. del Pozo et al. / Bioresource Technology 71 (2000) 143±149
Fig. 4. Evolution of e‚uent centrifuged COD during a shock load in the U-AFFR with a hydraulic retention time longer than 3 h.
the e‚uent centrifuged COD concentration because of
the shock load. E‚uent COD increased from 1550 to
2250 because of the Bv increase, which corresponded to
a decrease in the removal eciency from 72% to 58%.
After the shock load, eight days were required for the
reactor to recover its normal performance. However, no
signs of destabilization were detected, the reactor pH
was maintained around 7.4 by the bu€ering capacity of
the system and the alkalinity ratio (IA:PA) was always
below 0.56. A similar overload was applied to the
down¯ow reactor (D-AFFR). This showed the same
stability, but a shorter recovery time after the shock
because of the usual weekend break.
3.3. In¯uence of feed COD concentration
The reactors used were tubular-shaped, therefore
their performance could be in¯uenced not only by the
organic loading rate but also by the feed COD concen-
tration. In fact, Borja et al. (1995) detected VFA accu-
Table 3
Results obtained with di€erent feed COD concentrations and ®xed Bv
CODf (mg lÿ1 ) HRT (h) Bv (kg COD mÿ3 dÿ1 )
Average
2560 6 9.4
5080 15 8.0
11 300 31 8.3
147
mulation and an important removal eciency decrease
due to the inhibition of the digestion process, when
working with in¯uent abattoir wastewater COD con-
centrations of 29 000 mg lÿ1 in a down-¯ow ®xed-bed
reactor packed with clay rings.
In order to study the e€ect of feed organic matter
concentration, three di€erent feeds with average COD
concentrations of 2560, 5080 and 11 300 mg lÿ1 respec-
tively, were applied at a constant loading rate about 9 kg
COD mÿ3 dÿ1 . To achieve an increase in the concen-
tration of the poultry slaughterhouse wastewater,
chicken blood without clot, with an average COD of
75 000 mg lÿ1 , was added.
Table 3 shows the COD concentration inside the U-
AFFR reactor (CODe ) and the COD removal eciency
(% COD) for the di€erent feed concentrations (CODf )
used, the corresponding hydraulic retention times and
the organic loading rates. Under the operating condi-
tions of this experiment, feed concentrations ranging
from 2560 to 11 300 mg lÿ1 had no in¯uence on COD
CODe (mg lÿ1 ) % COD
r Average r
1 340 87 4
1 650 87 3
4 1200 89 1
148 R. del Pozo et al. / Bioresource Technology 71 (2000) 143±149
removal eciency, which remained constant at a ®xed
organic loading rate, regardless of the organic matter
concentration inside the reactor.
3.4. In¯uence of the hydraulic retention time
From the results discussed in the previous section and
summarized in Table 3, it was deduced that, at a ®xed
Bv around 9 kg COD mÿ3 dÿ1 , the process eciency did
not depend on HRT when it is longer than 6 h. But
when a considerably lower HRT (1.7 h) and a high or-
ganic loading rate (23±35 kg COD mÿ3 dÿ1 ) were ap-
plied, removal eciency decreased sharply to 38% and
biogas production fell quickly down to 30% of the
normal level. In spite of this, the alkalinity ratio (IA:PA)
was maintained at an acceptable value of 0.49 meaning
that methanogenic inhibition did not occur. However,
with this low HRT (1.7 h), a 50 kg COD mÿ3 dÿ1 shock
load produced a quick increase in the alkalinity ratio to
a dangerous value of 0.81. As indicated previously in the
section about transient organic overloads and in Fig. 4,
the application of shock loads when HRT was higher
than 2.8 h did not cause such destabilization, a fact
which shows the great in¯uence of very low HRT over
process stability.
Once operating conditions were relaxed by lowering
organic loading rate from 50 to 17 kg COD mÿ3 dÿ1 and
raising HRT from 1.7 to 5.5 h, the reactor reached its
usual biological activity in one week.
3.5. Intermittent operation
In order to study the reactor performance under real
slaughterhouse operating conditions, the in¯uence of
intermittent operation was determined, with weekly
breaks of 60 h over the weekend. The reactors were kept
at 35°C during the breaks.
The behavior of suspended biomass anaerobic reac-
tors (UASB) for dairy wastewater treatment under in-
termittent operation was studied by Carozzi (1993). He
found that weekend breaks cause biological unbalance
that drives to ¯oc disgregation and biomass loss. How-
ever, experience of anaerobic ®lters indicates that they
can tolerate frequent short breaks (Vincent, 1993).
In this work, satisfactory stability and removal e-
ciency were observed for more than a year of intermit-
tent operation. Although slight mineralization of the
suspended solids and the organic nitrogen occurred
during the break, endogenous decomposition of the
biomass during these periods was not signi®cant since
normal removal eciencies were reached within the ®rst
24 h after resuming feeding the reactor.
The stability observed under intermittent operation
was maintained even when the HRT was only 1.7 h and
also at a low temperature (20°C). For the highest or-
ganic loading rate fed (30 kg COD mÿ3 dÿ1 ), COD re-
moval eciency on the continuously operated U-AFFR
was 66%, while under intermittent operation (D-AFFR)
a slightly better removal of 71% was obtained, because
during the breaks VFA accumulation was mitigated.
3.6. In¯uence of recirculation
Recirculation can either increase or decrease removal
eciency of ®xed-®lm reactors depending on the extent
of the recirculation ¯ow and feed COD concentration
(Tritt, 1992). The presence of suspended biomass may
also be in¯uenced by super®cial velocity.
The reactors used in this work were operated with
recirculation ratios between 0 and 22, which implies
super®cial water velocities of up to 0.2 m hÿ1 , and within
an organic loading rate interval ranging from 7.0 to 15
kg COD mÿ3 dÿ1 . Each operating condition was main-
tained from 1 to 3 months. A statistical t-test showed,
with a con®dence level (a) of 0.05, that there was no
in¯uence of recirculation over COD removal. Conse-
quently, in the operation range studied, ®xed-®lm reac-
tors could be successfully operated without
recirculation, which implied minimum operating costs.
However, the relationship between recirculation and
suspended biomass inside the reactor should be con-
sidered in long-term studies.
3.7. In¯uence of temperature
In order to evaluate the feasibility of treating
slaughterhouse wastewater at the temperature it is pro-
duced; without any pre-heating, the anaerobic down¯ow
reactor was operated for 21 days at 20°C and a mod-
erate organic loading rate of 6.0 kg COD mÿ3 dÿ1 . The
eciencies obtained at 20°C and 35°C were compared
and it was observed that the COD removal eciency
decreased from 80% to 60%, which agrees with the re-
sults presented by other authors under similar condi-
tions (Viraraghavan and Kikkeri, 1990). In any case,
COD removal eciencies from 2160 to 860 mg lÿ1 jus-
tify wastewater pre-treatment without any external
heating.
4. Conclusions
The anaerobic ®xed-®lm reactor with non-random
support is a very appropriate system for pre-treatment of
wastewater with high organic load and high solids con-
centration, such as slaughterhouse e‚uents. It provides
satisfactory organic matter removal eciencies, even
when using high organic loading rates, under intermittent
operation, and with or without recirculation. Moreover,
under stressed operating conditions, such as shock loads,
very low hydraulic retention time or low temperature, the
reactor shows a quite stable performance.
 R. del Pozo et al. / Bioresource Technology 71 (2000) 143±149
Acknowledgements
The authors gratefully acknowledge ®nancial support
provided by the Universidad de Burgos, Junta de Castilla
y Leon (EB01/98) and Cooperativa Avõcola y Ganadera
de Burgos. R del Pozo is grateful to the Spanish MEC
for a fellowship.
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