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A comparative study of the industrial discharges effect on the anaerobic

treatment of domestic wastewater in both experimental and pilot-plant
scales
Ahlem Saddouda; Slim Abdelkafia; Fathi Alouia; Sami Sayadia
a
Laboratoire des Bioprocédés, Pôle d'Excellence Régional (PER, AUF), Centre de Biotechnologie de
Sfax, Sfax, Tunisia

Online publication date: 23 September 2010

To cite this Article Saddoud, Ahlem , Abdelkafi, Slim , Aloui, Fathi and Sayadi, Sami(2010) 'A comparative study of the
industrial discharges effect on the anaerobic treatment of domestic wastewater in both experimental and pilot-plant
scales', Environmental Technology, 31: 12, 1325 — 1333
To link to this Article: DOI: 10.1080/09593331003713701
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 Environmental Technology
Vol. 31, No. 12, November 2010, 1325–1333

A comparative study of the industrial discharges effect on the anaerobic treatment of

domestic wastewater in both experimental and pilot-plant scales
Ahlem Saddoud, Slim Abdelkafi, Fathi Aloui and Sami Sayadi*
Laboratoire des Bioprocédés, Pôle d’Excellence Régional (PER, AUF), Centre de Biotechnologie de Sfax, Sfax, Tunisia
(Received 15 December 2009; Accepted 18 February 2010 )
Taylor and Francis

10.1080/09593331003713701

The aim of this study was to compare the effect of industrial discharges on the anaerobic treatment of domestic
wastewater in both laboratory and pilot-plant scales at mesophilic conditions. The laboratory experiment results
have shown the low process efficiency of anaerobic treatment of DW by the use of an adapted or a non-adapted
methanogenic inoculum. These experiments performed in batch digesters were further confirmed by scaling up to
a pilot-plant anaerobic membrane bioreactor (MBR). The treatment inefficiency in both laboratory and pilot-plant
experiments could be related to the presence of toxic compounds due to the wastewater contamination by
industrial discharges. The toxic character of DW was proved by the phytotoxicity and microtoxicity tests. Indeed,
Downloaded By: [Monash University] At: 14:56 14 November 2010

the luminescence inhibition percentages started at an average of 21% in the morning and reached more than 84%
in the late afternoon. Moreover, the toxicity results have shown a direct relation with methanization results.
Indeed, when the average microtoxicity increased to 73%, the average germination index value and the
methanization efficiency expressed as the average methane percentage in the produced biogas decreased to 0% and
14.5%, respectively.
Keywords: domestic wastewater; industrial discharges; methanogenic inoculum; microtoxicity; phytotoxicity

Introduction Anaerobic digestion is an effective mean for waste-

In many Mediterranean countries, wastewater is not
always adequately treated, leading to deterioration of
existing freshwater resources and the Mediterranean Sea
[1,2]. The Mediterranean region is one of the areas in the
world most affected by water shortage. Moreover the
demand for water is expanding due to population
growth, increasing economic activities and expanding
areas of irrigated agriculture [3]. In addition, political,
economic and social boundary conditions changed
during the last years following new environmental regu-
lations. Political, social and economic outcome are
mainly promotion of treated wastewater reuse, reduction
of nuisance and protection of health and the environ-
ment [4]. Therefore, to ensure high treated wastewater
standards, there is a need to enhance the efficiency and
effectiveness of wastewater treatment and reuse.
As is well known, wastewater treatment processes
are inherently dynamic because of the large variations
in the wastewater flow rate, concentration and composi-
tion and their effects on the used treatment technologies
[5,6]. In fact, all over the world, several studies have
showed that anaerobic digestion technologies have been
successfully applied for treating domestic wastewater
[7,8].
water treatment which combines production of energy
with the decrease of the organic pollution of different
wastewaters [9]. This energy is mainly from a mixture
of methane and carbon dioxide, and if captured, is a fuel
resource for electricity generation or heat [10]. Anaero-
bic digestion is performed by different bacterial popula-
tions which are hydrolyzing bacteria, acid-forming,
acetate-forming and methane-forming bacteria which
act in sequence in the complex ecology. Methanogenesis
is performed by a class of Archaea aptly termed
methanogens [11].
Despite the large interest of the anaerobic digestion
processes, in some cases they cannot be used for the
treatment of problematic effluents, probably because of
their instability under some circumstances. Indeed,
anaerobic treatment limitations could be related to
numerous reasons which might include intrinsic limita-
tion that refers to the absence of biodegradative capac-
ity which can be due to the lack of cell uptake
mechanisms [12], lack of proper enzymes initiating
attack, lack of functionality [13,14], or unfavourable
thermodynamic reaction [15].
However, our recent studies [3,16] showed that the
anaerobic processes have limited efficiencies for the

*Corresponding author. Email: sami.sayadi@cbs.rnrt.tn

ISSN 0959-3330 print/ISSN 1479-487X online

© 2010 Taylor & Francis
DOI: 10.1080/09593331003713701
http://www.informaworld.com
 1326 A. Saddoud et al.
treatment of domestic wastewater contaminated by
industrial discharges. These discharges have a mineral
dominating character in the case of chemical industries
(such as surface treatment industries, i.e. chroming), an
organic dominating character in the case of detergent,
fish or olive mill factories and a mixed character for
textile and tannery industries [17,18].
Thus, the development of work aiming at the opti-
mization of the wastewater treatment process is a chief
stage to overcome the anaerobic treatment limitation.
The objectives of this work are:
● to reveal the effects of contaminated domestic
wastewater on the methanogenic inoculum
activity;
● to adapt the methanogenic inoculum for contami-
nated domestic wastewater treatment.
Downloaded By: [Monash University] At: 14:56 14 November 2010
Materials and methods
Domestic wastewater
Raw wastewater was sampled from the municipal
wastewater treatment plant (WWTP) localized at 5 km
in the south of Sfax, Tunisia. This WWTP is an aerated
lagoon process receiving industrial wastewaters. Its
treatment capacity is 24,000 m3 d−1. The wastewater had
an average chemical oxygen demand (COD) and
biological oxygen demand (BOD5) concentrations of
approximately 700 mg L−1 and 200 mg L−1, respec-
tively. The average characteristics of these effluents
were previously described [3]. For lab scale tests, the
raw wastewater samples were collected after the
primary treatment phase by an auto sampler in flasks,
then they were stored in the freezer. However, for pilot
scale tests the samples were stored at 4°C.
Experimental apparatus
The anaerobic batch digesters used in laboratory exper-
iments were constructed from 2000 mL flasks. At the
start of the experiments, each digester was filled with
400 mL of anaerobic inoculum and a defined volume of
substrate corresponding to the required COD concentra-
tion. These batch digesters had an equal final volume of
1000 mL. The anaerobic batch digesters were incubated
at 37 °C.
The experimental set-up used in the pilot-plant
experiment and consisted of an anaerobic membrane
bioreactor (MBR) which is constructed of Plexiglas
having a working volume of 50 litres. The temperature
was kept constant at 37 °C by circulating water through
the water jacket of the reactor. The reactor is coupled
via a multistage centrifugal pump Lowara SV805
(Frankfurt, Germany, 2–3 kW, Maximum flow rate =
10–12 m3 at 5–6 bars, with the frequency controlled by
a Stöber FBS/FDS) to a Technocon GmbH ultrafiltra-
tion system composed of a by a Stork membrane
module (Friesland BV, Germany) 3.048 m long. The
membrane, which was a Stork WFFX 0281 membrane,
had an area of 1 m2 area, and 100 kDa cut-off.
The adapted seed sludge used for batch digesters
and anaerobic MBR was obtained from a full-scale
anaerobic wastewater treatment plant. Moreover, the
non-adapted seed sludge used for batch digesters was
obtained from an anaerobic bioreactor treating olive
mill wastewater.
Analytical methods
Chemical oxygen demand (COD) was estimated using
the method described by Knechtel [19]. To determine
the gas composition, gas samples were collected with a
syringe from the tank of biogas and analyzed using a
gas chromatograph (Model IGC11 of DELSI, Delsi,
France) equipped with a thermal conductivity detector
and a concentric column (CTRI, Alltech, USA).
Column temperature was 60 °C. Helium was used as the
carrier gas at a flow rate of 35 mL min−1.
Volatile fatty acids (VFA) were analyzed using a gas
chromatograph (Shimadzu GC-9A, Kyoto, Japan)
equipped with a flame ionization detector (Shimadzu CR
6A, Kyoto, Japan). A Nukol capillary Silica column (30
m × 0.32 mm) was used. Oven, detector and injection
port temperatures were 100–150 °C, 250 °C and 250 °C,
respectively. The initial temperature of the column was
maintained at 100 °C for 2 minutes and then the temper-
ature was increased progressively (2 °C min−1) up to
150°C. One mL of mixed liquor sample was centrifuged
at 15,000 rpm for 15 min, supernatant was filtered
through a MilliPore® (Molsheim, France) membrane of
0.45 µm and 250 µL of filtrate were mixed with 10 µL
phosphoric acid (50%, v/v) and 1 µL was injected.
Microtoxicity
The microtoxicity tests consisted of inhibiting the biolu-
minescence of Vibrio fischeri LCK480 using the
LUMIStox system (Dr. Lange GmbH, Düsseldorf,
Germany) [20] in line with ISO Standard 11348-2 [21].
Since the luminescent bacterium requires marine condi-
tions, solid sodium chloride crystals were added to the
wastewater samples to a final concentration of 2% w/v.
The pH was adjusted to 7.0 ± 0.2. Wastewater or geomet-
ric dilutions were prepared with 2% sodium chloride
solution according to the Dr. Lange LUMIStox operating
manual. The rate of inhibition of the bioluminescence
was determined by mixing 0.5 mL of wastewater and 0.5
mL of luminescent bacterial suspension. After a 15-min
period of exposure at 15 °C, the decrease in the light
emission was measured. The toxicity of the wastewater
 Environmental Technology 1327

was expressed as the rate of inhibition of the biolumi- and 0.5 g COD in Batches 1, 2, 3, 4 and 5, respectively.
nescence relative to that of a non-contaminated control To introduce 0.5 g COD of domestic wastewater in Batch
sample. A positive control (7.5% NaCl) was included for 5, we use DW-concentrate (1.25-fold). In this data, two
each test. batch test digesters (Batch DW and Batch glucose) were
used. The first batch test digester fed with 100% of
glucose without domestic wastewater was used to inves-
Phytotoxicity tigate the effect of this effluent on glucose digestion by
Phytotoxicity was estimated by determining the germi- the exhausted methanogenic inoculum. The second
nation index (GI), as described by Zucconi et al. [22], batch test digester fed with 100% domestic wastewater
using Lepidium sativum seeds. without glucose was used to test the effect of the pres-
ence of an easily biodegradable co-substrate on the
digestion efficiency of domestic wastewater. These
Results and discussion batch digesters had an equal total volume of 1000 mL.
−1
Lg −COD
1
L −1);ofBatch byLa−1non L −1of −1 −1
). L −1);(0.2 DWL: −100%
1
(0.2 Lg−COD
1
L −1); Batch L −1)

As can be seen, the methane yield of glucose diges-

mixed
L
Figure
) ofwith
1.
glucose
Methane
33.3%
mixed
ofyield
domestic
withvariation
66.7%
wastewater
(0.4
during
g COD
(0.1
the co-digestion
) of domestic glucose
wastewater;
2: 50%
and domestic
ofBatch
glucose
5wastewater,
: (0.2
28.5%g COD
of glucose
) mixed
(0.2
adapted
gwith
CODmethanogenic
50% ) mixed
domestic
with
inoculum:
wastewater
71.5% Batch
of domestic
(0.2glucose
g COD
wastewater
: L100%); Batch
glucose
(0.5 3g: COD
(0.2
40%gof
LCOD
glucose g COD
Batch ) mixed
domestic
with 60%
wastewater
(0.3 g COD ) of domestic wastewater;
1: 66.7%Batch
of glucose
4: 33.3%
(0.2 g(0.2
CODg COD

Laboratory experiments
(1) Domestic wastewater methanization using a non-
adapted methanogenic inoculum
The activity of a non-adapted methanogenic inoculum
Downloaded By: [Monash University] At: 14:56 14 November 2010
tion is higher than the theoretical value. Such methane
production can be explained by the digestion of the
totality of glucose and a part of the anaerobic inoculum.
The comparison between methane yield values during

during the same sample treatment of domestic wastewa-
ter was evaluated by the use of an easily digested
substrate: glucose.
The experiments were carried out during two weeks.
Figure 1 shows the methane yield variation during the
co-digestion of glucose and domestic wastewater. Meth-
ane yield was calculated as the report produced methane/
theoretical methane potential of the substrate. Five
different batch digesters were fed by a glucose concen-
tration of 0.2 g COD L−1 mixed with different concen-
trations of raw domestic wastewater. These
concentrations were added to provide the desired COD
values varying from 0.3 to 0.7 g COD L−1 which corre-
spond to an added wastewater COD of 0.1, 0.2, 0.3, 0.4
the digestion of glucose and the co-digestion of glucose
and domestic wastewater shows that at low domestic
wastewater COD concentration, the methanogenic inoc-
ulum was able to degrade glucose. Also, the methane
yield increased from 45% in Batch 1 to 50% in Batch 2.
In this study, co-digestion was chosen because it
has become widely adopted as a method to improve
the performance of treatments [23]. In fact, the co-
digestion is a method where different types of wastes
are mixed and treated together and it offers many
advantages compared to the anaerobic digestion of one
substrate. However, Figure 1 shows a decrease of the
methane yield values with the increase of domestic
wastewater concentration introduced especially in

Figure 1. Methane yield variation during the co-digestion of glucose and domestic wastewater, by a non-adapted methanogenic
inoculum: Batch glucose: 100% glucose (0.2 g COD L−1); Batch DW: 100% domestic wastewater (0.2 g COD L−1); Batch 1:
66.7% of glucose (0.2 g COD L−1) mixed with 33.3% of domestic wastewater (0.1 g COD L −1); Batch 2: 50% of glucose (0.2 g
COD L−1) mixed with 50% of domestic wastewater (0.2 g COD L −1); Batch 3: 40% of glucose (0.2 g COD L−1) mixed with 60%
(0.3 g COD L−1) of domestic wastewater; Batch 4: 33.3% (0.2 g COD L−1) of glucose mixed with 66.7% (0.4 g COD L−1) of
domestic wastewater; Batch 5: 28.5% of glucose (0.2 g COD L−1) mixed with 71.5% of domestic wastewater (0.5 g COD L −1).
 1328
Batch digesters 3, 4 and 5. This decrease could not be
explained by glucose dilution and results given by the
Batch DW which was fed only with domestic waste-
water confirmed this deduction. Thus, taking into
account co-digestion results, we can assume that: first,
the biomass was unable to treat the domestic wastewa-
ter pollution domestic; and second, this effluent has an
inhibitor effect on glucose digestion by the non-
adapted methanogenic inoculum at a fraction of
domestic wastewater COD over than 50% of the total
COD concentration. Therefore, the metabolism was
deviated to the production of CO2 instead of CH4.
Figure 2 shows an example of CO2 and CH4 evolution
in Batch 5 and in Batch test. This deviation was
caused by the methanogenic disturbance created by
Downloaded By: [Monash University] At: 14:56 14 November 2010
Figure 2. Variation of CO2 (
) and CH4 (∆) percentage in Batch 5 (a) and in Batch test (b).
]qs[uf ]qs[u
A. Saddoud et al.
domestic wastewater presence at a fraction of 80% of
the total COD concentration.
In order to test the methanogenic inoculum ability in
Figure 2. Variation of CO 2 ([squf ]) and CH4 ([squ ]) percentage in Batch 5 (a) and in Batch test (b).
domestic wastewater digestion, nine samples of waste-
water collected at different days during a period of one
year, were used to feed nine different batch digesters
without glucose addition. Biogas production from 400
mg COD L−1 domestic wastewater digestion is given in
Figure 3.
As can be seen, almost 50% of all samples have a
Figure 3. Biogas production in 9 batch digesters fed by domestic wastewater without glucose.
very low biogas production. This may imply that the
methanogenic inoculum was enabling to digest these
effluents. The inability of methanogenic inoculum to
treat this effluent could be due to a possible presence of
low biodegradable substrates. To confirm this result,
 Environmental Technology 1329
Downloaded By: [Monash University] At: 14:56 14 November 2010

Figure 3. Biogas production in 9 batch digesters fed by domestic wastewater without glucose.

Batch 1 taken from the previous nine batch digesters
was fed only by glucose. After five days, 400 mg COD
L−1 of a new sample of domestic wastewater was
introduced in this batch (Figure 4). A high decrease of
the methanogenic inoculum activity caused by the intro-
duction of this effluent was shown. In addition, the limi-
tation of methanization process could be due to the
occurrence of toxic compounds coming from industrial
discharges in domestic wastewater treatment plant pipe-
lines. To investigate the effect of sampling time in
methanogenic inoculm activity, 15 samples of raw
domestic wastewater were collected during a day (6 h to
20 h) from domestic WWTP. Samples COD concentra-
tions and the evolution of methane percentage in

Figure 4. Effect of domestic wastewater on the methane (
) and biogas () production during the glucose digestion by a non-
adapted methanogenic inoculm.
 1330 A. Saddoud et al.
Downloaded By: [Monash University] At: 14:56 14 November 2010

Figure 5. COD evolution () of wastewater samples and variation of methane percentage (
) in the produced biogas.
]qs[u ]qs[uf

produced biogas are given in Figure 5. Results show
that methane percentage in the produced biogas could
be related to industrial and household activities. Indeed,
methane percentage started from 49.5% and decreased
considerably to reach at 19 h the least value of 14.5%.
This trend corroborated generally with the increase of
raw domestic wastewater COD concentration from the
morning to the afternoon. The VFA concentrations were
monitored in Batch 20 (Figure 6). Results showed that
total VFA concentrations did not exceed 200 mg L−1.
These concentrations were always below the inhibitory
limits in the batch digester. Therefore, the presence of
toxic compounds due to the wastewater contamination
by industrial discharges caused the decrease of the

Figure 6. Volatile fatty acid concentrations profile in the batch digesters Act (
), But (∆), ISB (×) , Val (▲), Total VFA ().
 Environmental Technology 1331

methanogenic inoculum activity. Indeed, domestic Phytotoxicity and microtoxicity tests were carried
wastewater toxicity could be related to industrial waste- out for monitoring the toxicity of these samples.
water discharges of the most problematic pollutants
occurring during the day, especially in the afternoon.
Thus, it can be concluded that only a part of the waste- (3) Domestic wastewater toxicity
water COD was biodegradable. The other part may be This part of the work attempted to determine the varia-
considered as a non-biodegradable or a recalcitrant tion of wastewater toxicity in function of the sampling
substrate. time. Therefore, samples used to investigate the effect
5.
6. Effect
Figure 4. COD
Volatile
evolution
offatty acid
([squ
domestic concentrations
]) of wastewater
wastewater on profilesamples
in the([squf
the methane batch
and variation
digesters
] ) and biogas
of
Actmethane
([squf But (∆),([squf
([squ percentage
]])), production
ISB (×])), in
during Val (▲
thethe
glucose
produced
), Total
digestion
VFA
biogas.
([squ
by a non
]). adapted methanogenic inoculm.

of time in adapted methanogenic inoculm activity were

also used in this section to evaluate their microtoxicity
(2) Domestic wastewater methanization by an adapted and phytotoxicity.
methanogenic inoculum
In order to test the methanogenic adapted inoculum
ability in domestic wastewater digestion, nine (a) Microtoxicity using LUMIStox test
samples of wastewater were used to feed nine differ- The results of toxicity tests showed that the percentage
ent batch digesters. Biogas production and methane of luminescence inhibition could be related to industrial
percentage evolution during domestic wastewater and household activities. Figure 8 shows a low percent-
digestion at a COD concentration of 400 mg L−1 is age of inhibition in the morning (until 10 h). The toxic-
Downloaded By: [Monash University] At: 14:56 14 November 2010

given in Figure 7. ity of the samples was high at 11, 12, 15, 18, 19 and
Figure 7 shows the same results obtained during
Figure 7. Biogas production ([squ ] ) and methane percentage ([squf ] ) during domestic wastewater digestion in the presence of an adapted inoculum.

20 h. The percentage of luminescence inhibition started

domestic wastewater digestion by a non-adapted inocu- at an average of 21% and reached up to more than 84%
lum. Thus, the low process efficiency of the anaerobic in the late afternoon.
wastewater treatment with an adapted or non-adapted Figure 8. Germination Index of Lepidium sativum ([squf ]) and toxicity profile using Vibrio fisheri ([squ ]) of wastewater samples taken from 6 a.m. to 08 p.m.

inoculum was proved. This process inefficiency could

be related to the wastewater composition. In fact, even (b) Phytotoxicity using Lepidium sativum
with an adapted inoculum, the wastewater methaniza- germination index test
tion was very low and impossible for some samples. The same samples of wastewater collected during a day
This may imply that the methanogenic inoculum was were tested for germination index of Lepidium sativum.
inhibited by the presence of toxic substrates resulting Results presented in Figure 8 showed that the GI varied
from the industrial discharges. from 0 to 22%. The phytotoxicity was lower in the

Figure 7. Biogas production () and methane percentage (
) during domestic wastewater digestion in the presence of an adapted ]qs[u ]qs[uf

inoculum.
 1332
Figure 8. Germination Index of Lepidium sativum (
) and toxicity profile using Vibrio fisheri () of wastewater samples taken
]qs[uf
from 6 h to 20 h.
Downloaded By: [Monash University] At: 14:56 14 November 2010
morning until 11 h while the GI decreased to 0% for the
more toxic samples of 12, 15, 16, 19 and 20 h. deter-
mined by the LUMIStox. A direct relation was
observed between the wastewater methanization
results, LUMIStox and the phytotoxicity tests. Indeed,
when the microtoxicity of the sample increased, the
germination index and the methanization efficiency
decreased.
Pilot-plant experiment
The laboratory experiments performed in batch digest-
ers were further verified by scaling up to a pilot-plant
anaerobic membrane reactor (MBR). The pilot-plant
was run with an organic loading rate from 0.8 to 1.2 g
COD L−1d−1. This work has been previously carried out
[24]. The results confirmed that the MBR proved to be
inefficient for the treatment of domestic wastewater
originated from an industrial region. The limitation of
the MBR treatment was shown through the considerable
variation of biogas production and methane percentage
in the biogas. This was due to the important variability
in the domestic wastewater composition and the exist-
ence of toxic compounds coming from the industrial
activities. Indeed, LUMIStox test and germination
index confirmed the toxic character of this effluent [24].
However, our previous work [25] has reported the
high efficiency of MBR in the anaerobic treatment of
domestic wastewater originated from a non-industrial
region. In this case, treated wastewater was of good
quality and was in line with World Health Organization
(WHO) guidelines for agricultural reuse. In fact, there
are significant differences between raw domestic waste-
water and contaminated domestic wastewater by indus-
trial discharges. Contaminated wastewater composition
A. Saddoud et al.
]qs[u
is complex due to the presence of a mix of fatty
compounds, proteins detergents, heavy metals and other
barely known compounds. These characteristics impose
limitations to the anaerobic process in respect of COD
removal efficiency, and also in terms of maximum
organic and hydraulic loading rates to be applied. These
limitations impose the need of integrated concepts for
domestic wastewater treatment.
The study by Aiyuk et al. [26] showed a high
efficiency of domestic wastewater treatment by an inte-
grated concept consisting of a coagulation/flocculation
pre-sedimentation step followed by treatment in an
upflow anaerobic sludge blanket reactor (UASB)
reactor.
Conclusion
The laboratory experiments have shown the low process
efficiency of the anaerobic treatment of domestic waste-
water by an adapted or non-adapted methanogenic inoc-
ulum. In addition, the pilot-scale experiments have
shown the same results.
The process inefficiency could be related to the
presence of toxic compounds due to the wastewater
contamination by industrial discharges which inhibited
both Lepidium sativum germination and Vibrio fischeri
luminescence. Thus, it may be advisable to choose an
urban locality for domestic wastewater collection and
avoid industrial contaminated localities.
Acknowledgements
We are grateful to the Office National d’Assainissement
(ONAS) for allowing us to take wastewater samples from the
wastewater treatment plant.
 Environmental Technology 1333

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