World Journal of Microbiology & Biotechnology (2005) 21:1509–1514 Ó Springer 2005

DOI 10.1007/s11274-005-7371-6

Anaerobic digestion of dairy manure in a hybrid reactor with biogas recirculation

Göksel N. Demirer1,* and Shulin Chen2

1
Dept. of Environmental Eng., Middle East Technical Univ., 06531, Ankara, Turkey
2
Dept. of Biological Systems Eng, Washington State Univ, 99163, Pullman, WA, USA
*Author for correspondence: Tel.: +90-312 210 58 67, Fax: +90-312 210 12 60, E-mail: goksel@metu.edu.tr

Received 17 March 2005; accepted 12 May 2005

Keywords: Anaerobic digestion, biogas recirculation, biomass immobilization, dairy manure, hybrid reactor

Summary

A novel anaerobic hybrid reactor (AHR) configuration incorporating floating support media for biomass immo-
bilization and biogas recirculation for enhanced mixing was used for anaerobic digestion of dairy manure. No
pretreatment or solid liquid separation was applied. The reactor was operated at high influent volatile solids (VS)
and organic loading rates (OLR) of up to 9.87% and 7.30 g VS/l day, respectively. After 149 days of continuous
operation the results revealed that a high amount (38.1 g VSS) of biomass was able to attach itself to the support
medium being used. The investigated AHR configuration achieved COD, BOD, TS, and VS removal efficiencies of
48–63, 64–78, 55–65, and 59–68%, respectively, at a hydraulic retention time (HRT) of 15 days. The corresponding
average methane production value obtained in this study was 0.191 l/g VS added.

Introduction
Anaerobic digestion is an established bioconversion
technology for high strength wastewater treatment.
Interest in using anaerobic digestion for manure man-
agement is rapidly growing as farmers and governments
are faced with mounting economic and environmental
concerns. The growing interest in the technology can be
attributed to anaerobic digestion’s potential for reduc-
ing pathogen levels and weed seeds, controlling odour,
improving fertilizer value, and producing value-added
products such as energy-rich methane; all of which can
help remedy or alleviate many of the economic and
environmental issues facing the growers and society.
Extensive research has been conducted and well docu-
mented on the feasibility of anaerobic treatment of farm
animal manure and its advantages, reactor types used,
performance, etc. (Lo & Liao 1985; Hobson & Wheatley
1993; Chynoweth et al. 1999, 2001; Demirer et al. 2000;
Demirer & Chen 2004; Güngör-Demirci & Demirer
2004). There are, though, several key areas of research
that must be pursued if anaerobic digestion technology
is to be made more economically advantageous. These
include investigating anaerobic digestion’s relatively low
digestion rates (or high retention time requirements) and
difficulties with biodegradation of lignocellulosic mate-
rial. Conventional high-rate anaerobic reactors such as
anaerobic filters, upflow anaerobic sludge blanket reac-
tors, etc. cannot effectively process wastes containing
more than 2–3% solids and are thus inappropriate for
use with animal manures which have much higher solids
content. Thus, demonstrating innovative anaerobic
process configurations that can process high solid ani-
mal waste at relatively short retention times will be an
innovative step towards achieving effective exploitation
of anaerobic digestion for animal manure.
Hybrid reactor configurations combine the positive
features of both suspended and attached growth systems
and constitute a promising option in anaerobic treat-
ment of different wastes. The anaerobic hybrid reactor
(AHR) which consists of a sludge bed in the lower part
and an anaerobic filter (AF) in the upper part combines
the advantages of upflow anaerobic sludge blanket
(UASB) and AF reactors (Suvajittanont & Chaiprasert
2003) while minimizing their limitations (Tilche & Vieira
1991). The application of AHRs in anaerobic treatment
include molasses (Boopathy & Tilche 1991), baker’s
yeast (Britz & van der Merwe 1993), coffee processing
(Bello-Mendoza & Castillo-Rivera 1998), slaughter-
house (Borja et al. 1995), pharmaceutical (Henry et al.
1996), ice-cream wastewater (Hawkes et al. 1995),
cheese whey (Malaspina et al.1996), phthalic waste (Tur
& Huang 1997), landfill leachate (Chang 1989), and
domestic sewage at low temperatures (Elmitwalli et al.
2002), as well as some studies focusing on improving
reactor design and operational parameters (Suvajitt-
anont & Chaiprasert 2003; Pender et al. 2004).
Increased biomass inventory and ability to process
high solid-containing wastes are two features of the
hybrid configuration which are significant for anaerobic
1510
digestion of animal manure. This is clear from the fact
that they have been successively used for digestion of food
wastes containing high solids concentrations (Hai-Lou
et al. 2002; Wang et al. 2002, 2003) as well as that of swine
(Lo et al. 1994) and dairy (Córdoba et al. 1995) opera-
tions. Even though it represents a viable option, hybrid
process configurations for anaerobic digestion of manure
have been the subject of a very limited number of inves-
tigations on poultry and swine manure (Yang et al. 1989,
1993) and none on dairy manure. Therefore, as a part of a
multifaceted and multi-disciplinary effort to address the
insufficiencies of current anaerobic digestion technology
for animal manure treatment, the objective of this study
was to investigate the performance of a novel AHR pro-
cess configuration, incorporating floating support med-
ium for biomass-immobilization and biogas recirculation
for enhanced mixing, in anaerobic digestion of dairy
manure.
Materials and methods
Dairy manure and anaerobic seed cultures
G.N. Demirer and Shulin Chen
litres of KONTAKTTM 565 support medium (NSW,
Roanoke, VI) with a diameter of 21.6 mm, thickness of
6.35 mm, dry weight of 121.9 g/l and specific surface area
of 564 m2/m3 was added to the AHR. A portion of the
biogas produced was recirculated through a cross-shaped
gas distributor with 1 mm openings by using a peristaltic
pump (Sigmamotor Inc., Middleport, NY). A wet-tip gas-
meter (Speece Tip, Nashville, TN) was connected to the
reactor to monitor the gas production. The AHR was
maintained at 36 °C (±2 °C) in a temperature-controlled
water bath. The performance of the reactor was moni-
tored by measuring biogas production, chemical oxygen
demand (COD), biological oxygen demand (BOD), total
solids (TS), volatile solids (VS), ammonia nitrogen (NH3–
N), total phosphorus (TP), and pH. Table 1 summarizes
the experimental plan used in this study. Before the onset
of daily feeding, the AHR was operated in a fed-batch
basis for 26 days. The purpose of this start-up period was
to acclimate the anaerobic cultures to the feed and start
biofilm formation on the support material. Continuous
operation was started on Day 27. AHR was operated
under seven different operational conditions which are
tabulated in Table 1.

Fresh manure was collected from the Dairy Center at

Washington State University in Pullman, WA and
stored at 4 °C prior to use. The composition of the raw
dairy manure is presented in Wen et al. (2004).
The mixed anaerobic culture used as seed was ob-
tained from the anaerobic lagoon of the Dairy Center at
Washington State University in Pullman, WA and
stored at 4 °C prior to use. The mixed anaerobic culture
was filtered through a screen of 0.0469 inch (1.19 mm)
mesh size and concentrated by settling for 2 h before
being used as inoculum. The volatile suspended solids
(VSS) concentration of the concentrated seed cultures
used was 4740 ± 240 mg/l.
Experimental set-up
The diameter, height, and the effective volume of the
AHR were 26 cm, 36.8 cm and 14.5 l, respectively. Three
Analytical methods
COD, BOD, TS, VS, and pH analysis were performed at
the WSU Water Quality Lab as described in Standard
Methods 5220D, 5210B, 2540B, 2540E, and 4500H,
respectively (APHA 1995). The content of CH4 in the
biogas was determined as follows. A known volume of
the headspace gas (V1) produced in a serum bottle used
in the biochemical methane production (BMP) experi-
ments was syringed out and injected into another serum
bottle which contained 20 g KOH/l. This serum bottle
was shaken manually for 3–4 min so that all the CO2
and H2S were absorbed in the concentrated KOH
solution. The volume of the remaining gas (V2), which
was 99.9% CH4, in the serum bottle was determined by
means of a syringe. The ratio of V2/V1 provided the
content of CH4 in the headspace (Ergüder et al. 2000).

Table 1. Operating conditions and performance data for the AHR.

Phase Operating conditions Performance data

VS in the COD HRT Recirculated Days Biogas Yield COD n BOD n TS n VS n NH3–N n TKN n TP n
Influent (%) loading Rate (days) Gas Volume operated (l biogas/g
(g/l day) (l/ day) VS added)

Start-up e e h 11.25 s m 1–26

1 1.66 0.87 20 11.25 s 27–47 0.226 ± 0.034 32–42 63–77 48–52 52–62 )10–8 12–16 20–24
2 4.98 2.61 20 33.75 s 48–69 0.299 ± 0.070 61–63 83–89 57–65 62–69 )11–11 14–36 19–35
3 8.30 4.35 20 33.75 s 70–78 0.234 ± 0.055 54–55 70–75 59 62 11 33 38
4 8.30 4.35 20 225.00 s 79–87 0.264 ± 0.025 64 73 47 47 )9 21 29
5 8.30 4.35 20 38.88 d 88–98 0.252 ± 0.011 53–56 69–78 55 55 18 14 19
6 9.87 6.89 15 38.88 d 99–125 0.293 ± 0.041 48–63 64–78 55–65 59–68 3–6 17–23 22–30
7 6.58 6.89 10 38.88 d 126–149 0.255 ± 0.032 30–58 57–73 26–50 30–44 6–10 8–9 8–17

e Variable; h fed-batch operation; m no biogas recirculation was applied if there was no biogas production observed the previous day during the
start-up period; s biogas recirculation rate was 750 ml/min; d biogas recirculation rate was 27 ml/min; n % removal.
Dairy manure digestion in hybrid reactor
Methane content of biogas was determined in six
duplicate samples and found to be 63.5 ± 3.7%.
Results
The performance of the reactor will be presented and
discussed for each operational phase (Phase 1–7). The
VS and COD loading rates applied, influent VS con-
centration, HRT, and gas recirculation data for each
operation phase are given in Table 1.
Phase 1
Following the start-up, continuous operation at an HRT
of 20 days was started with an organic loading rate
(OLR) of 0.92 g VS/l day (0.87 g COD/l day). The
biogas production rate and biogas yield aver
aged 3.02 ± 0.45 l/day and 0.226 ± 0.034 l biogas/
g VS added, respectively. The observed COD, BOD, TS,
VS, NH3–N, TKN, and TP removal efficiencies were 32–
42, 63–77, 48–52, 52–62, )10–8, 12–16, and 20–24%,
respectively. The observed COD removal efficiency was
more or less within the typical performance level of
anaerobic digesters treating dairy manure (Martin et al.
2003), while the observed VS removal was lower (Lo &
Liao 1985), similar to (Sánchez et al. 2000), and higher
(Dugba & Zhang 1999) than some of the studies in the
literature. The BOD data, however, cannot be compared
with literature data since most of the studies on anaer-
obic digestion of animal manure use COD and VS as the
parameters indicating the organic strength. The negative
value of ammonia nitrogen removal corresponds to an
increase in the reactor. This is an observation also re-
ported by other researchers (Cheng & Liu 2002; Martin
et al. 2003) and can be attributed to the anaerobic bio-
conversion of proteins contained in manure into amino
acids and then to ammonia. A possible explanation for
phosphorus removal is the accumulation in solids
remaining on the support media and settled in the reac-
tor. Supportingly Dugba et al. (1998) demonstrated that
the effluent from a sequencing batch reactor contained
less than 50% of the influent phosphorus. The balance of
the phosphorus was concentrated in the biosolids. The
relatively low treatment efficiencies observed for TKN
and TP are also expected, since anaerobic digesters are
known to reduce negligible amounts of nutrients.
Phase 2
The OLR was increased to 2.76 g VS/l day
(2.61 g COD/l day) after 21 days of operation at an
OLR of 0.92 g VS/l day. The corresponding VS percent
in the feed was 4.98. Furthermore, the volume of the
biogas recirculated in the system was increased from
11.25 to 33.75 l/day (Table 1). With this increase in
loading rate in Phase 2, the biogas yield did not reach its
steady-state level in Phase 1 immediately but increased
gradually. However, after five days of operation at
1511
Phase 2, the biogas yields observed exceeded
0.27 l biogas/g VS added, which was the maximum level
for Phase 1. After Day 54 the steady-state condition was
observed with respect to biogas production and the
average biogas yield for Phase 2 was determined as
0.299 ± 0.070 l biogas/g VS added. The corresponding
average biogas production rate was 11.97 ± 2.78 l/day.
The improved biogasification performance in Phase 2
relative to Phase 1 was also confirmed with higher
organics and solids removal efficiencies. The COD,
BOD, TS, and VS removal efficiencies were 61–63, 83–
89, 57–65, and 62–69%, respectively. Similar to Phase 1,
some of the average ammonia nitrogen concentrations
in the effluent were higher than that of the influent and
the removal efficiency ranged between )11 and 11%.
The TKN and TP removal efficiencies were a little
higher than Phase 1 and were between 14–36 and 19–
35%, respectively. The observed increase in COD, BOD,
and solids removal in the system in Phase 2 can be ex-
plained by acclimation of the anaerobic mixed cultures
to operating conditions, as well as an increase in gas
recirculation rate and higher influent organic concen-
trations. Even though it was not visually observed,
biofilm formation might also be another factor. It must
also be noted that not all of the solids removal observed
is necessarily associated with bioconversion, since it was
visually observed during the operation of the system
that support media in the reactor filtered out some of the
solid material in the influent.
Phase 3
In Phase 3, the OLR and influent VS concentration were
increased to 4.6 g VS/l day (4.35 g COD/l day) and
8.3%, respectively. The rest of the operating parameters
were kept the same (Table 1). Due to a higher OLR and
influent VS concentration, the biogas yield dropped
significantly from 0.299, the average value of Phase 2, to
0.222 l biogas/g VS added on Day 71. Starting with Day
71, the reactor was fed every other day until the end of
Phase 3 (Day 78) in order not to overload and inhibit
the anaerobic cultures. This countermeasure against
inhibition was successful, as depicted by the biogas
production and pH data. However, the biogas yield
values still remained low (0.192–0.214 l biogas/g VS).
The poor performance of the reactor at Phase 3 was also
reflected in the COD (54–55%), BOD (70–75%), TS
(59%), and VS (62%) removal data relative to Phase 2.
Phase 4
The only operational change made in Phase 4 was
increasing the biogas recirculation rate from 33.75 to
225 l/day (Table 1). The aim was to supply more mixing
through gas recirculation which could lead to improved
process performance. The increased biogas recirculation
rate resulted in an increase in biogas yield values to
0.264 ± 0.025 l biogas/g VS. However, its effect on
COD, BOD, TS, and VS removal efficiencies (64, 73, 47,
1512
and 47%, respectively) was not high enough to justify a
biogas recirculation rate increase by 6.7 times, which
would add significantly to operational costs.
Phase 5
After observing an insignificant increase in the process
performance by increasing the biogas recirculation rate
from 33.75 to 225 l/day in Phase 4, a different strategy was
adopted. Instead of further increasing the rate of biogas
recirculation from the Phase 2–3 level of 33.75 l/day, its
duration was extended 24 h and its rate was reduced to
27 ml/min (Table 1). The resulting volume of biogas re-
circulated in the system was 38.88 l/day, which corre-
sponded to a 15% increase from the Phase 2–3 level of
33.75 l/day. This practice did not lead to a better perfor-
mance in terms of biogas yield (0.252 ± 0.011 l biogas/
g VS). The average observed COD, BOD, TS, VS, NH3–
N, TKN, and TP removal efficiencies were 53–56, 69–78,
55, 55, 18, 14, and 19%, respectively.
Phase 6
The objective of this study was to investigate the per-
formance of a novel AHR process configuration on
anaerobic digestion of dairy manure; leading hopefully
to a superior performance relative to the existing sys-
tems. In particular, a significant achievement would be
the ability to maintain the existing performance while
achieving a certain HRT reduction which could lead to
reductions in the corresponding cost of the system
through volume reduction. Therefore, the HRT of the
reactor was reduced from 20 to 15 days in Phase 6. This
change increased the OLR and influent VS concentra-
tion to 7.30 g VS/l day (6.89 g COD/l day) and 9.87%,
respectively. It must be noted that the OLR of
7.30 g VS/l day adopted in this phase is higher than
most of the OLR values reported in literature for
anaerobic digesters treating dairy or cattle manure (Lo
& Liao 1985; Dugba & Zhang 1999; Harikishan & Sung
2003 Sung & Santha 2003) Similar to other phases where
an increase in OLR was practiced, a drop in the biogas
yield was observed. It decreased from the average level
of 0.252 ± 0.011 l biogas/g VS in Phase 5 to 0.201–
0.229 l biogas/g VS level on Days 100–102. However,
the biogas yield recovered from these low values rela-
tively fast ultimately reaching 0.300 l biogas/g VS level
on Day 111, until finally levelling off at 0.310–
0.338 l biogas/g VS for the rest of Phase 6 (Days 112–
125). The average biogas yield value for Phase 6 was
0.293 ± 0.041 l biogas/g VS which was significantly
higher than all the previous operational phases except
Phase 2. The COD, BOD, TS, and VS removal
efficiencies (48–63, 64–78, 55–65, and 59–68%, respec-
tively) observed in this phase were relatively high when
the operating HRT (15 days) is taken into consider-
ation. NH3–N, TKN and TP removal efficiencies ob-
served in Phase 6 (3–6, 17–23, 22–30%, respectively)
were not much different from the other phases.
G.N. Demirer and Shulin Chen
Even though the HRT was reduced to 15 days in
Phase 6, the operation was very stable in terms of biogas
production and yield after the first 10 days (Days 110–
125). The reduction in the HRT of the system was not
conducted before Phase 6 (or after 99 days of operation)
when biofilm formation was clearly observed from the
transparent lid of the reactor. Thus, it could be specu-
lated on the basis of this visual observation that the
increase in the biomass concentration through biofilm
formation led to a more stable operation with higher
performance. Because it required opening the lid of the
reactor, which would negatively affect the anaerobic
conditions in the reactor, the biofilm formation was not
quantified in Phase 6. However, after operation of the
reactor was terminated, support media samples were
quantified for attached biomass, as will be discussed
below.
Phase 7
After acquiring better process performance at a reduced
HRT of 15 days in Phase 6, a further decrease of HRT
to 10 days was investigated in Phase 7. Since only the
effect of further reduction in the HRT was to be inves-
tigated, the OLR value from Phase 6 (7.30 g VS/l day or
6.89 g COD/l day) was kept constant. This needed the
influent VS concentration to be reduced to 6.58%
(Table 1). Further reduction in the HRT did not lead to
a sudden change in the performance of the system. The
biogas yield varied between 0.258 and 0.320 l biogas/
g VS for Days 126–137, however a gradual but steady
decrease in both biogas production and yield was ob-
served until Day 143 at which point the biogas yield
stabilized between 0.215 and 0.236 l biogas/g VS. The
average biogas yield for Phase 7 was determined as
0.255 ± 0.032 l biogas/g VS which was considerably
lower than for Phase 6 (0.293 ± 0.041 l biogas/g VS).
The low COD, BOD, TS and VS removals (30–58, 57–
73, 26–50, and 30–44, respectively) were consistent with
the biogas yield data. When the sharp increase in the
effluent VS and TS concentrations is considered along
with the lower but continuous biogas production rates
in the reactor, a gradual wash-out from the reactor
could easily be concluded. This explains the observed
poor performance of the reactor in Phase 7. The average
biogas yields and ranges of COD, BOD, TS, VS, NH3–
N, TKN, and TP removal efficiencies for each phase are
depicted in Table 1.
Attached biomass quantification
At the end of the operation (Day 149), the attached
biomass on support medium was measured as described
in Punal et al. (1999). The results indicate that each
medium had an average attached biomass of
50.8 ± 7.4 mg VSS which corresponds to a total at-
tached biomass of 38.1 g VSS or 12.7 g VSS/l within the
upper zone of the AHR which contained the support
media. It should be noted that the measured total
Dairy manure digestion in hybrid reactor
attached biomass (38.1 g) was present in the 3 l support
media zone, while the rest of the AHR volume (11.5 l)
contained suspended biomass. The biofilm formation
takes extended periods in anaerobic reactors (Punal
et al. 1999; Elmitwalli et al. 2002; Suvajittanont &
Chaiprasert 2003), due to the low growth rates of the
anaerobes. This, in turn, may explain the observed
higher performance levels in Phase 6.
Discussion
When the significantly high loading rate (6.89 g COD/
l day) and low HRT value (15 days) used in this study,
and their obvious cost reduction impact, are taken into
consideration, the average methane production value
obtained in this study (0.191 l/ g VS added) represents a
superior performance over the studies reported in the
literature. This enhanced process performance is
thought to be mainly due to better mixing through
biogas recirculation and biomass immobilization on the
support media as supported by the findings of Keshtkar
et al. (2003) and Suvajittanont & Chaiprasert (2003).
Therefore, based on the results of this study, it can be
concluded that an AHR configuration incorporating
floating support media for biomass immobilization and
biogas recirculation for enhanced mixing can be suc-
cessfully used for the anaerobic digestion of dairy
manure at high concentrations and loading rates. It
must be noted that anaerobic digestion has to be fol-
lowed by a polishing treatment step (aerobic or chemi-
cal) for discharge into a receiving environment.
This study generated a preliminary understanding of
the performance of the proposed AHR configuration
under different set of operating conditions. The
encouraging performance observed calls for further
investigation of the significant operating variables such
as biogas recirculation, intensity of mixing, type and
amount of support media, temperature, etc.
Acknowledgments
The authors express sincere appreciation to Mr Craig
Frear for his valuable comments and careful proof-
reading.
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