Bioresource Technology 98 (2007) 130134

Anaerobic co-digestion of algal sludge and waste paper

to produce methane
Hong-Wei Yen
a

a,

, David E. Brune

Department of Chemical Engineering, Tunghai University, Taiwan, ROC

Department of Biosystems Engineering, Clemson University, SC, USA

Received 6 June 2005; received in revised form 11 November 2005; accepted 12 November 2005
Available online 4 January 2006

Abstract
The unbalanced nutrients of algal sludge (low C/N ratio) were regarded as an important limitation factor to anaerobic digestion process. Adding high carbon content of waste paper in algal sludge feedstock to have a balanced C/N ratio was undertaken in this study. The
results showed adding 50% (based on volatile solid) of waste paper in algal sludge feedstock increased the methane production rate to
1170 75 ml/l day, as compared to 573 28 ml/l day of algal sludge digestion alone, both operated at 4 g VS/l day, 35 C and 10 days
HRT. The maximum methane production rate of 1607 17 ml/l day was observed at a combined 5 g VS/l day loading rate with 60% (VS
based) of paper adding in algal sludge feedstock. Results suggested an optimum C/N ratio for co-digestion of algal sludge and waste
paper was in the range of 2025/1.
2005 Elsevier Ltd. All rights reserved.
Keywords: Anaerobic digestion; Co-digestion; C/N; Methane; Algal sludge

1. Introduction
The recently patented Partitioned Aquaculture System (PAS) has been proven to be able to increase aquaculture production capacity by improving pond waste nutrient
treatment (Brune et al., 2001). In the PAS, algae play a dual
role, as both an oxygen supplier to the Wsh and as a waste
nutrient cleaner. The capacity of Wsh production in the PAS
has been demonstrated to be four times greater than that of
conventional aquaculture and also at the same time the
wastewater discharges are signiWcantly decreased. However
concentrated algal sludge production from the PAS must
be continually removed from the ponds to control algal
density and water column respiration. The large amount of
algal sludge represents a potential source of fuel and recovered N and P fertilizer (Mulbry et al., 2005). Anaerobic
digestion of algal sludge produced from the PAS can not
*
Corresponding author. Tel.: +886 4 235 90 262x209; fax: +886 4 235 90
009.
E-mail address: hwyen@thu.edu.tw (H.-W. Yen).

0960-8524/$ - see front matter 2005 Elsevier Ltd. All rights reserved.
doi:10.1016/j.biortech.2005.11.010

only decrease the amount of waste to be handled, but also

yields methane to oVset the energy needs of the PAS achieving a sustainable aquaculture production system.
The solar energy stored in the algal biomass in results of
the photosynthesis reaction could be released as methane
through the anaerobic digestion. This concept was originally proposed over 40 years ago in a paper by Oswald and
Golueke (1960), describing integrated processes of largescale raceway pond cultivation of microalgae and wastewater treatment, followed by fermentation of algal biomass to
methane fuel. Due to the recalcitrant of algal sludge to biodegradation, Chen and Oswald (1998) found the heat pretreatment of algal sludge at 100 C for 8 h could improve
the eYciency of methane fermentation a maximum at 33%.
However, the improvement on the methane energy produced would not be economically competitive to the energy
lost on the heat pretreatment.
Except for the resistant to biodegradation, the low C/N
ratio of algal sludge is also a serious problem to the anaerobic digestion. Although, an optimum C/N range in feedstock for the anaerobic digestion is still debatable in the

H.-W. Yen, D.E. Brune / Bioresource Technology 98 (2007) 130134

literature, 20/130/1 is a most acceptable range (Parkin and

Owen, 1986). The C/N ratio in algal sludge is about 6/1,
which is too low for the digestion. Low C/N ratio feedstock
could result in high total ammonia nitrogen (TAN)
released and high volatile fatty acids (VFAs) accumulated
in the digester. The TAN and VFAs both are important
intermediates and potential inhibitors in the anaerobic
digestion process (Parkin and Owen, 1986). High concentration of TAN and VFAs in the digester would decrease
the methanogen activity and further accumulation could
fail the anaerobic digestion. One method to avoid excessive
ammonia accumulation is to adjust low feedstock C/N
ratios by adding high carbon content materials, thereby
improving the digestion performance. This practice has
been used for co-digestion of sewage sludge and municipal
solid waste (MSW) (Sosnowaki et al., 2003). Most MSW
consists of paper material (including oYce and newspaper),
which has a C/N ratio ranging from 173/1 to greater than
1000/1 while typical sewage sludge has a C/N ratio ranging
from 6/1 to 16/1 (Stroot et al., 2001). Co-digestion of cattle
manure slurry with fruit, vegetable wastes and chicken
manures is another example of successful blending of high
C/N and low C/N feedstocks to improve digester performance (Callaghan et al., 2002). Co-digestion of sisal pulp
and Wsh wastes had shown a 5994% increased in the methane production yield as compared to sisal pulp and Wsh
wastes digestion alone (Mshandete et al., 2004). The beneWts of co-digestion include: dilution of potentially toxic
ammonia, allowing for increased loading rate and
improved biogas yield (Sosnowaki et al., 2003).
The purpose of this work was to assess the possibility of
co-digestion of algal sludge and high carbon content of
waste paper at diVerent fraction to produce methane and
evaluate the waste paper adding eVects on the methane production.
2. Methods
2.1. Anaerobic digestion experiments
Bench-top anaerobic digesters with gas storage tanks
were fabricated from 25 mm Plexiglas sheet and contained
with a temperature-controlled water bath at 35 1 C.
Digesters were connected to gas storage tanks and gas sampling ports using silicone tube. The digester and gas storage
tank measured 12.5 14 30 cm and 16.5 15.2 30 cm,
respectively (working volume, 4l). Semi-continuously feeding type was adopted and digesters were fed once per day
following the removal of the same volume of eZuent. Hand
mixing of the digester was performed with a permanently
installed mixing rod prior to eZuent removal and immediately after loading. The digestion was operated at 10 days
HRT. The pH was 6.5 and was controlled (based on the
samples pH) by adding 5 N sodium hydroxide solution, if
necessary. The daily biogas production was recorded by
measurement of water displacement. Daily samples were
stored at 4 C for analysis. The digesters were operated for

131

20 days and then assumed to be in steady-state (variation in

daily biogas production was within 10% of average production). Furthermore, a NR ratio ( D TKNin/TKNout)
between 0.95 and 1.05 was used as a measure of steady-state
conditions (Cobbs and Hill, 1990).
2.2. Characteristics of substrates
Algal sludge and waste paper were used as feeding substrates in this investigation. Algal sludge was harvested
from Partition Aquaculture System, Clemson University.
The species of algae in the sludge would change by seasons,
but most of the species would be Scenedesmus spp. and
Chlorella spp. Waste paper was collected from recycle bins
at the Clemson Computing and Information Technology
labs (DCIT). This paper was used in laser printer with one
side printed mostly and was cut by shredder or scissor
into 0.5 1 cm pieces before mixing with algal sludge for
feeding.
2.3. Analytical methods
Biogas was collected from the digester gas sampling
ports with gas sampler tube. The tubes were allowed to
sample for 1015 s before shutting the outlet valve to insure
that the sample was representative of digester gas. The biogas composition was determined using a gas chromatograph (SRI-8610C, SRI instruments) with a thermal
conductivity detector (TCD). A 0.5 ml of gas sample was
injected into the chromatograph with a column temperature of 45 C, using helium as the carrier gas. The sample
gas concentration was compared to a standard gas mixture
consisting of 40% methane and 60% carbon dioxide, and
60% methane and 40% carbon dioxide. Glucose-release rate
from conversion of carbomethycellulose (CMC) by cellulase during a predetermined time period was used as measurement method of cellulase activity. Cellulase activity is
expressed as mg of glucose-released per ml of sample per
minute at 38 C. Soluble total ammonia nitrogen (TAN)
was measured using an ammonia sensor (ORION-720A).
Volatile fatty acids (VFAs) and total kjeldahl nitrogen
(TKN) were determined following standard methods
(APHA, 1995).
3. Results and discussion
Total carbon and kjeldahl nitrogen was 10,500 mg/l and
2000 mg/l, and 406 mg/g and 182 mg/kg in algal sludge and
waste paper, respectively. The C/N ratios were about 5.3/1
in algal sludge and 2000/1 in waste paper.
3.1. Algal sludge digestion
Digesters fed with algal sludge alone at 2, 4 and 6 g VS/
l day loading rate were undertaken to investigate the digestibility of algal sludge and following methane production
rate. As seen in Table 1, methane production rates

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H.-W. Yen, D.E. Brune / Bioresource Technology 98 (2007) 130134

Table 1
Methane production rates, VFAs, and TAN (SD) in algal sludge digestion operated at diVerent loading rates and 10 days HRT
Loading rate (g VS/l day)

CH4 (ml/l day)

CO2 (ml/l day)

VFAs (mg/l)

TAN (mg/l)

2
4
6

180 8
573 28
818 96

72 5
258 15
384 44

1305 148
4978 635
5862 1769

589 86
837 65
998 29

increased in proportion to increases in loading rate. The

methane production rate was 180 8 ml/l day at 2 g VS/
l day loading rate, and increased to a maximum methane
production rate of 818 96 ml/l day at 6 g VS/l day loading
rate. Both VFAs and TAN concentrations increased with
increasing loading rate to 5682 1769 mg/l and 998
29 mg/l at 6 g VS/l day loading rate. It was expected that the
VFAs and TAN concentration eventually would achieve
the toxicity concentration, if the loading rate kept increasing. Methane production yields were 90, 143 and 136
CH4 ml/g VS introduced at loading rates of 2, 4 and 6 g VS/
l day, respectively. Methane production yields of 100140
CH4 ml/g VS introduced at 10 days HRT acquired in this
investigation were lower than the yield of 260 ml methane/g
VS introduced at 30 days HRT acquired by Oswald and
Golueke (1960). Apparently, a longer HRT would be detrimental to have a higher methane production yield. However, HRT increase would decrease the loading rate, and
the lower loading rate would have the less methane production rate, as seen in Table 1. The reactor capital cost was
considered as the major investment in the anaerobic digestion (Rivard, 1993). Therefore, introducing shorter HRT
and higher loading rate to minimize the reactor volume
required could be a good choice for having a economical
competitive digestion process.
3.2. Co-digestion of algal sludge and waste paper
at a loading rate of 4 g VS/l day
The results of co-digestion of algal sludge and waste
paper at diVerent blending fractions at 4 g VS/l day loading rate were shown in Table 2. Five ml of trace elements
per day and 350 mg-N as ammonium chloride were added
each day in the digester of waste paper to prevent from
the nutrients limitation. The waste paper fractions in the
co-digestion with algal sludge were on the volatile solid
basis. Co-digestion of algal sludge and paper blended at

4 g VS/l day loading rate, with paper addition at 50% of

VS, led to signiWcant increases in the methane production
rate to 1170 75 ml/l day; or two-fold higher than that
observed in algal sludge digestion alone. According to the
observed methane production rates in Table 2, neither
algal sludge alone nor waste paper alone was an optimum
substrate for anaerobic digestion. Digester TAN levels
decreased with the increased feedstock C/N ratio to the
minimum of 65 10 mg/l at 75% of waste paper fraction.
Digesters fed with 50% algal sludge and 50% paper
yielded relatively low VFA levels of 3912 1290 mg/l
compared to 4978 635 mg/l during algal sludge digestion
alone. With the increase of C/N ratio to 36.4/1 at a 75%
paper and 25% algal sludge, the digester performance
turned to be unstable. Possible reasons for this observed
instability could be (1) toxicity of high VFAs at
10,848 713 mg/l, and (2) low TAN concentrations of
65 10 mg/l.
A balanced C/N ratio in feedstock was likely to be beneWcial to the methanogen activity and resulted in VFAs concentration decreased by more VFAs converted to methane.
However, at 75% paper fraction in feedstock, the C/N ratio
of 36.4/1 was possibly too high for the anaerobic digestion,
which led to low TAN levels of 65 10 mg/l observed.
McCarty (1964) reported that 50200 mg/l of TAN was
required for growth of anaerobic microorganism. Therefore, low TAN levels of 65 10 mg/l resulting from high C/N
ratio feedstock was likely limiting growth of the microbial
population in these digesters.
Vinzant et al. (1990) reported that paper digestion
under aerobic environment was more eYcient than the
digestion under anaerobic environment. Also, 12 days of
HRT in anaerobic digestion of paper was the minimum
required for a stable digester operation (Vinzant et al.,
1990). Therefore, it was believed 10 days HRT used in this
study was too short for the paper digestion alone. In the
batch of paper digestion alone, even if the C/N was

Table 2
Methane production rates, VFAs and TAN concentrations (SD) in co-digestion of algal sludge and waste paper at loading rate 4 g VS/l day (with diVerent waste paper fractions) and 10 days HRT
Feedstock

C/N

CH4 (ml/l day)

CO2 (ml/l day)

VFAs (mg/l)

TAN (mg/l)

Algal sludge
(25%) of waste paper + algal sludge
(50%) of waste paper + algal sludge
(75%) of waste paper + algal sludgea
(100%) of waste paperb

6.7
11.8
18.0
36.4
21.5

573 28
968 73
1170 75
317 114
452 36

258 15
556 37
769 49
277 82
278 22

4978 635
3480 679
3912 1290
10,848 713
7350 1259

837 65
697 51
524 24
65 10
629 51

a
b

Digestion performance is not at stable situation.

Adding trace element and NH4Cl 87.5 mg-N/l day as the nitrogen source.

H.-W. Yen, D.E. Brune / Bioresource Technology 98 (2007) 130134

3.3. Cellulase activity in the co-digestion of algal sludge

and waste paper at 4 g VS/l day
Co-digestion of waste paper and algal sludge could
eYciently balance feedstock carbon and nitrogen and a
balanced C/N ratio of feedstock was likely to beneWt the
methane production rate. The increase of cellulase activity
resulting from paper addition was another possible reason
for explaining increased methane production rate. Cellulase are inducible enzymes which are synthesized and
mostly secreted into the environment by microorganisms
during their growth on cellulosic materials, oligo and dimmer sugars, including some of their derivates (Busto et al.,
1996). Cellulose hydrolysis is considered the rate-limiting
step in digesters fed with a high cellulosic content feedstock. It was suggested that paper addition to the digester
might induce cellulase excretion by bacteria such as Clostridium themocellum (Suto and Tomita, 2001). In fact, in
the digesters fed paper and algal sludge blends at 4 g VS/
l day loading rate, cellulase activity increased, resulting in a
positive impact on the methane production rate (Fig. 1). At
75% paper fraction, elevated VFA concentrations suppressed cellulase activity to 1.26 0.14 mg/l min. The highest level of cellulase activity (3.02 0.09 mg/l min) was
during paper digestion alone. However, the methane production rate was only 452 36 ml/l day as compared to
573 28 ml/l day for algal sludge digestion alone. These
results suggested that breakdown of algal biomass contributed some key components to the improvement of methanogenic activity. Therefore, a low methane production rate
was observed in the digester fed paper alone, even if there
were no nutrients limitation and had high cellulase activity
in the digester (see Table 3).

3.5
Cellulase activity
3

Glu. released mg/l m

adjusted to 21.5/1 by adding ammonic chloride, it still had

a low methane production rate of 452 36 ml/l day. One
possible explanation for this was that algal biomass was
not only playing a nitrogen source supplier in this codigestion process, but also supplied nutrients to the
digester microXora after the degradation of algal biomass.
This might be able to explain why the co-digestion of algal
sludge and waste paper had a higher methane production
rate than the paper digestion alone, even if the C/N ratio of
digester fed paper alone was adjusted to 21.5/1 by adding
ammonium chloride.

133

2.5
2
1.5
1
0.5
0

25%

50%

75%

100%

Paper fraction in feedstock

Fig. 1. Cellulase activity in digesters fed algal sludge and waste paper at a
combined loading rate of 4 g VS/l day and 10 days HRT.

3.4. Co-digestion of waste paper and a Wxed amount

of algal sludge
The eVects of varying waste paper loading (0, 1, 2, 3 and
4 g VS/l day) blended with a Wxed amount of algal sludge
loading at 2 g VS/l day on co-digestion performance were
investigated. Digesters fed at a Wxed algal sludge loading
rate of 2 g VS/l day and paper loading rates of 0, 1, 2, 3 and
4 g VS/l day yielded a maximum methane production rate
1607 17 ml/l day at a combined loading rate of 5 g VS/
l day (C/N D 22.6/1). This decreased to 856 40 ml/l day at
a combined loading of 6 g VS/l day (C/N D 27.2/1). With
the increase of loading rate, the VFAs increased and TAN
levels decreased. Since nitrogen source in this co-digestion
was came from the breakdown of algal sludge, at a Wxed
algal sludge loading (2 g VS/l day), TAN levels decreased
with the increased paper loading. That meant the waste
paper in the combined feedstock turned to be the main carbon source instead of algal sludge. Results suggested that
at a loading rate of 6 g VS/l day, the paper fraction in feedstock achieved 67% (C/N D 27.2) was too high for the
anaerobic digestion process, which led to lower methane
production rate than 5 g VS/l day loading rate. C/N ratio
between 20 and 25/1 was optimum (Fig. 2) for the anaerobic co-digestion of algal sludge and waste paper at 10 days
HRT and 35 C.

Table 3
Methane and carbon dioxide production rates, VFAs and TAN concentrations (SD) in digesters at 2 g VS/l day of algal sludge loading and varying loadings of waste paper (0, 1, 2, 3, 4 g VS/l day) and 10 days HRT
Combined loading (g VS/l day)

C/N

CH4 (ml/l day)

CO2 (ml/l day)

VFAs (mg/l)

TAN (mg/l)

2
3
4
5
6

6.7
13.3
18.0
22.6
27.2

180 34
823 16
1170 75
1607 17
856 40

72 5
397 14
769 49
1080 21
561 23

1305 147
3780 458
3912 1290
5220 855
6228 685

589 86
541 2
524 24
396 8
175 7

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H.-W. Yen, D.E. Brune / Bioresource Technology 98 (2007) 130134

350
algal sludge + paper

Methane yield, CH4 ml/g VS in

300

250

200

150

100
paper + NH4

50

0
0

10

15

20

25

30

35

40

C/N
Fig. 2. Methane yield vs. C/N ratio in digesters with algal sludge and paper
loading at combined loading rates of 2, 3, 4, 5 and 6 g VS/l day and 10 days
HRT.

4. Conclusions
Co-digestion of algal sludge and waste paper was useful
and oVered two beneWts: (1) a balance of C/N ratio. The
optimized C/N range for the co-digestion was 2025/1; (2)
increase in cellulase activity. The increase in cellulase activity might be helpful in the biodegradation of algal sludge,
which could provide nutrients in the digester, which Wnally
would improve methane production rate.
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