Bioresource Technology 98 (2007) 130–134

Anaerobic co-digestion of algal sludge and waste paper

to produce methane
a,¤ b
Hong-Wei Yen , David E. Brune
a
Department of Chemical Engineering, Tunghai University, Taiwan, ROC
b
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 pro-
cess. 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 20–25/1.
© 2005 Elsevier Ltd. All rights reserved.

Keywords: Anaerobic digestion; Co-digestion; C/N; Methane; Algal sludge

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

The recently patented “Partitioned Aquaculture Sys-
tem” (PAS) has been proven to be able to increase aquacul-
ture 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 recov-
ered 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).
yields methane to oVset the energy needs of the PAS achiev-
ing 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 origi-
nally proposed over 40 years ago in a paper by Oswald and
Golueke (1960), describing integrated processes of large-
scale raceway pond cultivation of microalgae and wastewa-
ter treatment, followed by fermentation of algal biomass to
methane fuel. Due to the recalcitrant of algal sludge to bio-
degradation, Chen and Oswald (1998) found the heat pre-
treatment 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 pro-
duced 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 anaero-
bic digestion. Although, an optimum C/N range in feed-
stock for the anaerobic digestion is still debatable in the

0960-8524/$ - see front matter © 2005 Elsevier Ltd. All rights reserved.
doi:10.1016/j.biortech.2005.11.010
 H.-W. Yen, D.E. Brune / Bioresource Technology 98 (2007) 130–134
literature, 20/1–30/1 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 concen-
tration 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 perfor-
mance (Callaghan et al., 2002). Co-digestion of sisal pulp
and Wsh wastes had shown a 59–94% increased in the meth-
ane production yield as compared to sisal pulp and Wsh
wastes digestion alone (Mshandete et al., 2004). The bene-
Wts 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 pro-
duction.
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 sam-
pling 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, 4–l). Semi-continuously feed-
ing 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 immedi-
ately after loading. The digestion was operated at 10 days
HRT. The pH was 6.5 and was controlled (based on the
sample’s 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 pro-
duction). 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 sub-
strates 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 10–15 s before shutting the outlet valve to insure
that the sample was representative of digester gas. The bio-
gas composition was determined using a gas chromato-
graph (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 tempera-
ture 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 cellu-
lase during a predetermined time period was used as mea-
surement 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 digest-
ibility of algal sludge and following methane production
rate. As seen in Table 1, methane production rates
132 H.-W. Yen, D.E. Brune / Bioresource Technology 98 (2007) 130–134
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)
2 180 § 8
4 573 § 28
6 818 § 96
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 increas-
ing. 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 100–140
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 detri-
mental to have a higher methane production yield. How-
ever, HRT increase would decrease the loading rate, and
the lower loading rate would have the less methane produc-
tion rate, as seen in Table 1. The reactor capital cost was
considered as the major investment in the anaerobic diges-
tion (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 load-
ing 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
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 diVer-
ent waste paper fractions) and 10 days HRT
Feedstock C/N CH4 (ml/l day)
Algal sludge 6.7 573 § 28
(25%) of waste paper + algal sludge 11.8 968 § 73
(50%) of waste paper + algal sludge 18.0 1170 § 75
(75%) of waste paper + algal sludgea 36.4 317 § 114
(100%) of waste paperb 21.5 452 § 36
a
Digestion performance is not at stable situation.
b
Adding trace element and NH4Cl 87.5 mg-N/l day as the nitrogen source.
CO2 (ml/l day) VFAs (mg/l) TAN (mg/l)
72 § 5 1305 § 148 589 § 86
258 § 15 4978 § 635 837 § 65
384 § 44 5862 § 1769 998 § 29
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 bene-
Wcial to the methanogen activity and resulted in VFAs con-
centration 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 50–200 mg/l of TAN was
required for growth of anaerobic microorganism. There-
fore, 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
CO2 (ml/l day) VFAs (mg/l) TAN (mg/l)
258 § 15 4978 § 635 837 § 65
556 § 37 3480 § 679 697 § 51
769 § 49 3912 § 1290 524 § 24
277 § 82 10,848 § 713 65 § 10
278 § 22 7350 § 1259 629 § 51
 H.-W. Yen, D.E. Brune / Bioresource Technology 98 (2007) 130–134 133

adjusted to 21.5/1 by adding ammonic chloride, it still had 3.5

a low methane production rate of 452 § 36 ml/l day. One Cellulase activity
possible explanation for this was that algal biomass was 3
not only playing a nitrogen source supplier in this co-

Glu. released mg/l m

digestion process, but also supplied nutrients to the 2.5
digester microXora after the degradation of algal biomass.
This might be able to explain why the co-digestion of algal 2
sludge and waste paper had a higher methane production
rate than the paper digestion alone, even if the C/N ratio of 1.5
digester fed paper alone was adjusted to 21.5/1 by adding
ammonium chloride. 1

3.3. Cellulase activity in the co-digestion of algal sludge 0.5

0 25% 50% 75% 100%
and waste paper at 4 g VS/l day
Paper fraction in feedstock

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. Cellu-
lase are inducible enzymes which are synthesized and
mostly secreted into the environment by microorganisms
during their growth on cellulosic materials, oligo and dim-
mer 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 feed-
stock. It was suggested that paper addition to the digester
might induce cellulase excretion by bacteria such as Clos-
tridium 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 sup-
pressed cellulase activity to 1.26 § 0.14 mg/l min. The high-
est level of cellulase activity (3.02 § 0.09 mg/l min) was
during paper digestion alone. However, the methane pro-
duction 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 contri-
buted some key components to the improvement of metha-
nogenic 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).
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 car-
bon source instead of algal sludge. Results suggested that
at a loading rate of 6 g VS/l day, the paper fraction in feed-
stock 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 anaero-
bic 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 load-
ings 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 6.7 180 § 34 72 § 5 1305 § 147 589 § 86
3 13.3 823 § 16 397 § 14 3780 § 458 541 § 2
4 18.0 1170 § 75 769 § 49 3912 § 1290 524 § 24
5 22.6 1607 § 17 1080 § 21 5220 § 855 396 § 8
6 27.2 856 § 40 561 § 23 6228 § 685 175 § 7
134 H.-W. Yen, D.E. Brune / Bioresource Technology 98 (2007) 130–134

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100
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