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Available at www.sciencedirect.com journal homepage: www.elsevier.com/locate/watres Thermophilic anaerobic digestion of source-sorted organic fraction of household municipal solid waste: Start-up procedure for continuously stirred tank reactor Irini Angelidaki a, , Xingxing Chen a , Junbo Cui a , Prasad Kaparaju a , Lars Ellegaard b a Institute of Environment and Resources -DTU, Building 113, Technical University of Denmark, DK-2800, Kgs. Lyngby, Denmark b Burmeister and Wain Scandinavian Contractor A/S, Gydevang 35, DK-3450 Allerød, Denmark article info Article history: Received 15 February 2006 Received in revised form 11 May 2006 Accepted 17 May 2006 Keywords: Anaerobic digestion Batch Full-scale Lab-scale Methane yield SS-OFMSW Start-up Thermophilic ABSTRACT Two feeding strategies for start-up of continuously stirred tank reactors (CSTR) treating source-sorted organic fraction of household municipal solid waste (SS-OFMSW) at 55 1C were evaluated. Two reactors were started up separately with a limited amount of initial inoculum (i.e. 10% of the final volume of 3.5 l) and operated in a fed batch mode until the reactors were filled (30 days). A reference reactor was filled up with 3.5 l of inoculum and fed at a constant rate (11.4 g volatile solids (VS)/d). Loading at progressively increasing rate (from 1.7 to 15 gVS/d), as calculated based on an activated biomass concept, showed superior process performance compared to a fixed feed rate (5.7 gVS/d). Methane yield of 0.32 m 3 /kg VS was produced during the start-up in reactor filled at progressively increasing rate and was comparable to the reference reactor. On the contrary, significant inhibition due to volatile fatty acid (VFA) build-up, mainly due to butyrate, was noticed in the reactor filled at constant rate. Thus, low initial and progressive increasing inoculum loading rate could be used as a strategy for a successful start-up of CSTR treating SS-OFMSW as it allowed a gradual acclimation of the biomass. Lab-scale results were further reaffirmed from the start-up of a full-scale plant (7000 m 3 total capacity) which was supplied with inoculum corresponding to approx. 16% of final volume and operated in a fed batch mode until the reactors were filled (58 days). Stable biogas production with low VFA (o3 g/L; based on titration method) were noticed during the start-up period when fed at progressively increasing rate. Thus, a controlled and reliable start-up procedure was found essential, which could allow rapid process stabilization and time to focus on other technical aspects of plant operation. In addition, the influence of substrate to inoculum amount (1.5–30% TS) and temperature (5–65 1C) on anaerobic degradation and methane production of SS-OFMSW was investigated in batch assays as a protocol for start-up procedure. & 2006 Elsevier Ltd. All rights reserved. 1. Introduction Anaerobic digestion of organic fraction of municipal solid waste (OFMSW) has increasingly being studied and applied in Europe, especially after the introduction of source-sorted (SS) collection (De Baere, 2000). Currently, more than one million tonnes of organic wastes (wet weight) per year are digested in dedicated industrial plants worldwide (Bolzonella et al., 2003a). Earlier, full-scale plants treating OFMSW were oper- ated only at mesophilic temperatures (Cecchi et al., 1993) and ARTICLE IN PRESS 0043-1354/$ - see front matter & 2006 Elsevier Ltd. All rights reserved. doi:10.1016/j.watres.2006.05.015 Corresponding author. Tel.: +45 45251429; fax: +45 45932850. E-mail address: [email protected] (I. Angelidaki). WATER RESEARCH 40 (2006) 2621– 2628

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ARTICLE IN PRESS

Available at www.sciencedirect.com

WAT E R R E S E A R C H 4 0 ( 2 0 0 6 ) 2 6 2 1 – 2 6 2 8

0043-1354/$ - see frodoi:10.1016/j.watres

�Corresponding auE-mail address:

journal homepage: www.elsevier.com/locate/watres

Thermophilic anaerobic digestion of source-sorted organicfraction of household municipal solid waste: Start-upprocedure for continuously stirred tank reactor

Irini Angelidakia,�, Xingxing Chena, Junbo Cuia, Prasad Kaparajua, Lars Ellegaardb

aInstitute of Environment and Resources -DTU, Building 113, Technical University of Denmark, DK-2800, Kgs. Lyngby, DenmarkbBurmeister and Wain Scandinavian Contractor A/S, Gydevang 35, DK-3450 Allerød, Denmark

a r t i c l e i n f o

Article history:

Received 15 February 2006

Received in revised form

11 May 2006

Accepted 17 May 2006

Keywords:

Anaerobic digestion

Batch

Full-scale

Lab-scale

Methane yield

SS-OFMSW

Start-up

Thermophilic

nt matter & 2006 Elsevie.2006.05.015

thor. Tel.: +45 45251429; [email protected] (I. Angelida

A B S T R A C T

Two feeding strategies for start-up of continuously stirred tank reactors (CSTR) treating

source-sorted organic fraction of household municipal solid waste (SS-OFMSW) at 55 1C

were evaluated. Two reactors were started up separately with a limited amount of initial

inoculum (i.e. 10% of the final volume of 3.5 l) and operated in a fed batch mode until the

reactors were filled (30 days). A reference reactor was filled up with 3.5 l of inoculum and

fed at a constant rate (11.4 g volatile solids (VS)/d). Loading at progressively increasing rate

(from 1.7 to 15 gVS/d), as calculated based on an activated biomass concept, showed

superior process performance compared to a fixed feed rate (5.7 gVS/d). Methane yield of

0.32 m3/kg VS was produced during the start-up in reactor filled at progressively increasing

rate and was comparable to the reference reactor. On the contrary, significant inhibition

due to volatile fatty acid (VFA) build-up, mainly due to butyrate, was noticed in the reactor

filled at constant rate. Thus, low initial and progressive increasing inoculum loading rate

could be used as a strategy for a successful start-up of CSTR treating SS-OFMSW as it

allowed a gradual acclimation of the biomass. Lab-scale results were further reaffirmed

from the start-up of a full-scale plant (7000 m3 total capacity) which was supplied with

inoculum corresponding to approx. 16% of final volume and operated in a fed batch mode

until the reactors were filled (58 days). Stable biogas production with low VFA (o3 g/L; based

on titration method) were noticed during the start-up period when fed at progressively

increasing rate. Thus, a controlled and reliable start-up procedure was found essential,

which could allow rapid process stabilization and time to focus on other technical aspects

of plant operation. In addition, the influence of substrate to inoculum amount (1.5–30% TS)

and temperature (5–65 1C) on anaerobic degradation and methane production of SS-OFMSW

was investigated in batch assays as a protocol for start-up procedure.

& 2006 Elsevier Ltd. All rights reserved.

1. Introduction

Anaerobic digestion of organic fraction of municipal solid

waste (OFMSW) has increasingly being studied and applied in

Europe, especially after the introduction of source-sorted (SS)

r Ltd. All rights reserved.

x: +45 45932850.ki).

collection (De Baere, 2000). Currently, more than one million

tonnes of organic wastes (wet weight) per year are digested in

dedicated industrial plants worldwide (Bolzonella et al.,

2003a). Earlier, full-scale plants treating OFMSW were oper-

ated only at mesophilic temperatures (Cecchi et al., 1993) and

ARTICLE IN PRESS

Table 1 – The characteristics of substrate (SS-OFMSW)before dilution

Parameter SS-OFMSW

TS (%w/w) 30

VS (%w/w) 24.4

pH 4.6

TotalVFA (g/kg-waste) 27.3

Acetate (g/kg-waste) 22.2

Propionate (g/kg-waste) 3.6

Iso-butyrate (g/kg-waste) 0.34

Butyrate (g/kg-waste) 0.50

Iso-valerate (g/kg-waste) 0.46

Valerate (g/kg-waste) 0.24

Total N (g-N/kg-waste) 6.5

NH4+-N (g-N/kg-waste) 1.5

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reported to show good operational performance (Buhr and

Andrews, 1977). But with successful demonstration of anae-

robic treatment of OFMSW at thermophilic temperature both

in lab-scale (Wellinger et al., 1992) and full-scale (Cozzolino

et al., 1992), the thermophilic anaerobic process has gained

increased attention primarily due to its capacity for higher

loading rate and greater volumetric gas production potential

(Cecchi et al., 1991). In addition, the thermophilic process has

been established as a reliable and accepted mode of

fermentation as claimed by several plant operators (Mata-

Alvarez et al., 2000) because of enhanced hydrolysis, in-

creased organic solids destruction and better pathogen

control (Ahn and Forster, 2000; Lund et al., 1995). Despite

these advantages, application of the thermophilic process in

the past has been limited mainly due to difficulty in process

control, poor effluent quality and poor process stability

related to chronically high propionate concentrations (Kugel-

man and Guida, 1989). The latter problems were often

encountered in connection with or shortly after start-up.

Start-up is therefore an important step in establishing a

proper microbial community in thermophilic as well as other

biological treatment processes. Poor start-up in biological

treatment systems can lead to prolonged period of acclima-

tion (Wu et al., 2001) and ineffective removal of organic

matter (Griffin et al., 1998). Several researchers have dis-

cussed the general start-up procedure for biogas plants

treating solid waste (Bolzonella et al., 2003a, b; Fernandez et

al., 2001; Lepisto and Rintala, 1995). These studies reported

that successful start-up was related to a number of factors

e.g., seed sludge source, initial loading rate, hydraulic

retention time (HRT) and/or solids retention time (Fang and

Lau, 1996). Some of these studies also included a start-up

strategy such as use of seed material from plants treating

similar waste, gradual process acclimatization from meso-

philic to thermophilic and/or gradual organic load increase

and withholding feeding temporally after inoculation (e.g.,

Bolzonella et al., 2003a; Cecchi et al., 1993; Fernandez et al.,

2001; Fang and Lau, 1996; Van Lier et al., 1993).

For a successful start-up and to reach the design load

quickly, strategies such as the amounts of inoculum and the

initiation of the feeding should be designed to avoid

accumulation of anaerobic degradation intermediate pro-

ducts such as propionic acid and other volatile fatty acids

(VFA’s) and hydrogen, which could inhibit methanogenesis

and acetogenesis (Lepisto and Rintala, 1995). Several re-

searchers have considered and demonstrated that thermo-

philic anaerobic processes can be readily started with small

amounts of inocula from a thermophilic process or with any

mesophilic sludge having methanogenic activity (Cecchi

et al., 1993; Fang and Lau, 1996; Lepisto and Rintala, 1995;

Van Lier et al., 1992, 1993). However, inability to acquire large

quantities of thermophilic inocula as seed material and long

and unstable start-up of the thermophilic process when

inoculated with mesophilic seed in practice, has led to poor

reputation with respect to process stability. In this study,

feeding strategies for starting thermophilic reactors with only

an initial amount of 10% of the final volume were evaluated. A

new strategy was designed based on an ‘‘activated biomass’’

concept (see later section for definition of concept), where

loading is increased in relation to the content of ‘‘activated

biomass’’ in the reactor. In this way daily loading is applied in

proportion to the gradual build-up of microbial degradation

capacity, avoiding the risk of accumulation of VFA and other

intermediates. A reference reactor started with 100% inocu-

lum, was fed with the same amount of fresh substrate every

day. Furthermore, the influence of substrate to inoculum

amount (1.5–30% total solids, TS) and temperature (5–65 1C) on

anaerobic degradation and methane production of SS-

OFMSW were investigated in batch assays.

2. Material and methods

2.1. Origin of substrate and inoculum

SS-OFMSW from a full-scale biogas plant (Vaarst–Fjellerad

plant; Denmark) was used as substrate. At the biogas plant,

substrate was shredded and sorted in multiple steps to yield a

purified digester feed that was homogenized, enriched in

biodegradable organics and devoid of large pieces or plastic

stringers. In the lab, substrate was further blended and

shredded to smaller pieces to prevent clogging. The prepared

feed was then stored in plastic bags of approx. 2 kg at �20 1C

until use. Characteristics of the substrate used are presented

in Table 1.

Thermophilic digested manure from a full-scale biogas

plant (Vegger plant, Denmark) treating manure together with

industrial waste and OFMSW was used as inoculum.

2.2. Batch experiments

The effect of substrate to inoculum ratios by varying substrate

concentration on anaerobic degradation of SS-OFMSW was

performed in 550 ml glass serum bottles. To each assay, 150 ml

of inoculum and 100 ml of a mixture of SS-OFMSW and tap

water corresponding to different initial TS contents of 0%,

1.5%, 3%, 4.5%, 6%, 7.5%, 9% and 30% were added. Anaerobic

conditions in the assays were established by flushing the

headspace with nitrogen gas for 3 min. Assays were sealed

immediately with butyl rubber stoppers and aluminum

crimps. All experiments were conducted in duplicate and

incubated statically at 55 1C in a temperature controlled

ARTICLE IN PRESS

Table 2 – Percentage conversion of fresh feed to ‘‘activebiomass’’ based on feed activation concept for loading ata progressive increasing rate

Feed activation concept

Days ai Conversion (%)

1 a1 25

2 a2 20

3 a3 20

4 a4 15

5 a5 10

6 a6 5

7 a7 5

WAT E R R E S E A R C H 40 (2006) 2621– 2628 2623

chamber. Assays containing inoculum and water were used

as controls. Methane produced from inoculum alone was

subtracted from the methane produced in other serum

bottles for comparison of methane production from fresh

substrate. Experiments were terminated after 40 days when

no significant gas production was noticed.

In another series of batch assays, the effect of temperature

on methane production potential of SS-OFMSW with thermo-

philic inoculum was evaluated. To each assays, 150 ml of

inoculum and 100 ml of SS-OFMSW diluted to 4.5% TS with

tap water were added as described above and incubated at

temperatures of 5, 15, 20, 30, 37, 45, 55 and 65 1C. The

experiments were conducted in duplicate and incubated for

75 days.

Total 100

Note: ai is the conversion percentage of substrate during the first 7

days after feeding.

2.3. Start-up procedure of CSTR treating SS-OFMSW

Three separate 4.5 litres (l) CSTRs referred to as R1, R2 and R3

with liquid volume of 3.5 l were operated at 55 1C with heating

water pumped through the reactor water jackets. Reactor

contents were mixed at 40 s/min by mechanical mixers. On

day 1, 3.5 l of inoculum (i.e. 100% inoculation) was introduced

into reactor R1 and operated continuously with 15 d HRT

(reference reactor). In reactors, R2 and R3, 350 ml of inoculum

(i.e. 10% inoculation relative to final volume) was introduced

prior to substrate addition. The substrate was diluted with tap

water in weight ratio of 1:4 to achieve 6% TS (w/w) feed. The

reactors were fed twice every day by peristaltic pumps.

Feeding strategies were loading at a progressive increasing

rate (from 1.7 to 15 gVS/d) in R2 and at constant rate of 5.7 gVS/

d in R3 while the reference reactor R1 was fed at a constant

rate of 11.4 gVS/d. Thus both R2 and R3 were filled up to final

volume 3.5 l in 30 d.

2.4. Calculation of loading rate for reactor fed at aprogressive increasing rate (R2)

Loading rate for R2 was calculated based on a feed ‘‘activation

concept’’. This concept is based on constant loading rate (10%)

of calculated ‘‘activated biomass’’ content. Initial inoculum

added to the reactor is considered as ‘‘activated biomass’’ and

subsequent addition of fresh biomass is assumed to be

converted gradually to ‘‘activated biomass’’ over a period of

7 days after feeding at a conversion rate as shown in Table 2.

During the first day, 25% of the feed is assumed converted into

‘‘activated biomass’’ and on the second day 20% and so on. In

a continuous loading situation the amount of ‘‘activated

biomass’’ on any given day is the amount of ‘‘activated

biomass’’ present on the previous day plus 25% of the fresh

feed from the previous day plus 20% of the fresh feed 2 days

before etc. This concept can easily be implemented in a work

sheet starting with initial inoculum at day 0. In a formula the

principle can be expressed as

ABn ¼ ABn�1 þX7

i¼1

Fn�iai,

where AB ¼ activated biomass; F ¼ daily feed; n ¼ day num-

ber; ai ¼ conversion percentage day i ¼ 1–7 (Table 2).

This concept results roughly in an exponential increasing

loading designed to follow the gradual formation of microbial

capacity. Relative loading rate can be adjusted during the

start-up phase based on VFA analysis.

2.5. Start-up of full-scale plant based on the ‘‘activatedbiomass’’ concept

The ‘‘activated biomass’’ start-up concept has also been

applied in the start-up of a full-scale plant (Lemvig Biogas

plant, Denmark). Lemvig biogas plant is a centralised biogas

plant treating manure in co-digestion with various types of

organic industrial waste. The plant consists of 3 reactors with

a total digester volume of approx. 7000 m3. An initial amount

of 1100 m3 of thermophilic inoculum (i.e. approx. 16% of final

volume) was supplied to the plant during the first 10 days

after the initial start-up from another thermophilic biogas

plants in the region. Feed batch operation was continued for

58 days, after which all 3 reactors were full and effluent

pumping was started. Intended daily loading was 5% of the

calculated ‘‘activated biomass’’ content according to a con-

cept as described above. However, actual influent could not

always follow the intended feed rate due to technical

disturbances (pump break downs etc.). The three reactors

were for practical reasons (mixing and heating) started-up

sequentially. R1 was the first to be filled and when full, R1

content was divided into R1+R2 and finally the R1+R2 contents

were divided into R1+R3. Therefore, the VFA analyses for R2

and R3 were available only after some time.

2.6. Analytical methods, laboratory experiments

pH was measured with Metrohm 744 pH meter immediately

after each sampling. Methane content in the biogas was

analysed using a gas chromatography (GC-14A) equipped with

TC detection. The biogas produced in CSTRs was measured

through gas meters (Angelidaki et al., 1992). VFA concentra-

tions were determined using GC5890-series II equipped with

flame ionisation detection (FID). Total nitrogen (sum of

ARTICLE IN PRESS

WAT E R R E S E A R C H 4 0 ( 2 0 0 6 ) 2 6 2 1 – 2 6 2 82624

ammonia nitrogen and organic bound nitrogen), TS and VS

were determined according to Standard Methods (APHA,

1998).

3. Results and discussion

3.1. Effect of substrate-inoculum ratios by varyingsubstrate concentration (1.5–30% TS) on anaerobic digestionof SS-OFMSW at 55 1C

Table 3 shows the specific methane yields for SS-OFMSW

incubated with inoculum at varying substrate concentration.

After an initial lag phase (5–7 d), a slow initial methane

production was noticed in all assays (data not shown). This

phase was followed by a sharp increase in the methane

production due to degradation of accumulated intermediate

compounds. A similar pattern was reported in a study using

spent brewery grain and anaerobic sludge incubated at 37 1C

in batches at different inoculum-substrate ratios of 7%, 13%

and 20% (Fernandez et al., 2001).

Specific methane yields after 40 days of incubation, in

general, increased with decrease in SS-OFMSW solids content

(Table 3). The maximum specific methane yield of 0.41 m3/

kgVS was obtained at 1.5% and 3% TS, while 63–83% of this

value were achieved at 4.5–9% TS. At 30% TS, the process was

completely inhibited due to overloading, with a methane

production lower than that obtained from control. The slow

rates and prolonged methane production at TS contents

higher than 1.5% indicate inhibition. Jungersen and Ahring

(1994) also reported that degradation of liquefied manure was

most efficient at relatively low concentrations, whereas a

significant inhibition occurred at higher concentrations. The

high specific methane yield at 1.5% TS was attributed to the

fact that complete anaerobic degradation could be achieved

in those assays with high inoculum to substrate ratio (low

substrate concentration with constant inoculum volume),

without VFA accumulation reaching inhibiting levels. The

present results thus suggest that during start-up of a

conventional digester like CSTR, where the acid and methane

producing phases are together, it is necessary to optimise for

Table 3 – Specific methane yields of varying TS content ofSS-OFMSW after 40 days at 55 1C

TS content (%) Specificmethane yield(m3/kgVS added

waste)

Relativemethane yieldcompared to

1.5%TS

1.5 0.41 (1.3) 100

3 0.41 (1.5) 99.7

4.5 0.35 (1.5) 83.9

6 0.26 (1.6) 63.5

7.5 0.26 (1.7) 62.2

9 0.30 (2) 73.5

30 Inhibition Inhibition

Note: Values in parentheses are standard deviation.

a low initial amount of fresh organic material relative to

inoculum amount. In the present study, the initial amount of

fresh organic material was varied by dilution of the fresh

substrate. The same ratio between fresh organic material and

inoculum could also be achieved by applying a reduced

amount of undiluted fresh material, however without the

potential benefit of general dilution of potential inhibitors

and minor accumulation of intermediates. Upon successful

start-up, a gradual increase in TS of fresh feed can be

attempted.

3.2. Effect of temperature (5–65 1C) on anaerobic digestionof SS-OFMSW

The specific methane yields from assays incubated with

thermophilic inoculum at various temperatures for 75 days

are shown in Table 4. Methane production rates (data not

shown) and the final yields were generally higher at

temperature closer to that of original inoculum. The low

methane yields at 30 1C and below were probably due to the

fact that the bacterial culture was not adapted to the low

temperatures and a long adaptation period is required for

such a non-adapted thermophilic culture (Nozhevnikova

et al., 2002).

The present results demonstrate the importance of obtain-

ing inoculum adapted to the same temperature level as

intended for the process to be started-up or at least to seek

inoculum adapted to a temperature within 710 1C of the new

process. Otherwise long start-up periods are required for the

development and retention of high concentrations of active

and well balanced biomass inside the reactor. This is

especially important for thermophilic processes because only

9% thermophiles and 1% obligate thermophiles were reported

to be present in mesophilic sludge (Chen, 1983). The fact that

the highest specific methane production was achieve at 45 1C,

somewhat below the process temperature at the source of

inoculum could be due to temperature dependent ammonia

concentration.

Table 4 – Mean-specific methane yields from SS-OFMSW(4.5% TS) incubated in batch assays at different tem-peratures (5–65 1C)

Temperature(1C)

Specificmethane (m3/

kgVS)

Relativemethane

compared to1.5%TS

5 0.001 (0) 0.2

15 0.003 (0.01) 0.5

20 0.13 (0.5) 22.3

30 0.30 (1.2) 51.6

45 0.58 (2.2) 100

55 0.42 (1.5) 71.8

65 0.52 (0.46) 85.5

Note: Values in parentheses are standard deviation.

ARTICLE IN PRESS

R1 (100% inoculum)

0

3

6

9

VF

A c

ompo

nent

(g/L

)

R2 (Progressive increasing loading rate)

0

3

6

9

VF

A c

ompo

nent

(g/L

)

R3 (Constant loading rate)

0

3

6

9

0 10 20 30 40

Time (day)

VF

A c

ompo

nent

(g/L

)

Fig. 2 – Monitoring the VFA components during the start-up

of anaerobic digestion SS-OFMSW at 55 1C in CSTR: J,

acetate; K, propionate; &, butyrate; ’, iso-butyrate; n,

valerate; m, iso-valerate.

0

150

300

450M

etha

ne y

ield

(mL/

gVS

)

0

5

10

15

0 10 20 30 40

Time (day)

Load

ing

rate

(gV

S/d

)

Fig. 1 – Loading rate (below) and methane production (above)

during the start-up of thermophilic anaerobic digestion of

SS-OFMSW (6% TS) in CSTR: R1 (J, constant rate with 10%

inoculum), R2 (&, progressive increasing rate with 10%

inoculum) and R3 (n, constant loading rate with 100%

inoculum).

WAT E R R E S E A R C H 40 (2006) 2621– 2628 2625

3.3. Start-up procedure of continuously stirred tankreactor treating SS-OFMSW

Start-up process performance of the CSTRs is illustrated in

Fig. 1. Reactor R2 responded well to the loading strategy of a

progressively increasing rate, although R2 was inoculated at

10% volume compared to that of R1. The initial low-feeding

strategy approach in R2 has apparently (as intended) allowed

a gradual balance between the various degradation steps to

develop sufficient microbial degradation capacity thereby

avoiding any possible VFA build-up. Total VFA content in R2

dropped steadily from 5.2 to 1.3 g/L within 12 d and remained

under 0.2 g/L thereafter (Fig. 2). These VFA values indicate a

stable start-up process and were in accord with those values

of 0.4 g/L reported earlier during start-up process of mechani-

cally sorted (MS) OFMSW (Cecchi et al., 1993). Correspond-

ingly, the specific methane production in R2 increased at first

slowly (first 3 days) producing only 0.06 m3/kgVS, increasing

sharply on day 4 (0.11 m3/kgVS) and then steadily thereafter

to produce 0.32 m3/kgVS at the end (Fig. 1). A similar

development in biogas production was noticed in the

reference reactor R1 fed at constant rate. Specific methane

yields of 0.31–0.32 m3/kgVS produced in R1 and R2 respectively

were comparable to other results obtained during thermo-

philic treatment of MS-OFMSW. For instance, Pavan et al.

(2000) reported a methane yields of 0.32 m3/kgVS obtained

during thermophilic semi-dry digestion of OFMSW (25%TS)

fed at loading rate of 9.7 kgVS/m3 d. Feeding R3 (also inocu-

lated with 10% of final volume as R2) at a constant rate on the

contrary resulted in a sharp increase in VFA concentration,

especially on day 7 (6.7 g/L), and resulted in a very low specific

methane production (0.10 m3/kgVS) which even remained low

throughout the start-up period (Fig. 2). This low specific

methane yield in R3 was apparently due to process inhibition

caused by VFA accumulation (Bolzonella et al., 2003a). After

day 15, methane production in R3 increased steadily to reach

0.20 m3/kgVS in the end but remained much lower than that

produced in R1 and R2.

Among the measured VFA components, acetate, propionate

and n-butyrate were the predominant acids. Specific distribu-

tion of VFAs in the three reactors indicated that with time,

acetate in R1 decreased gradually (from 6.8 to o0.06 g/L)

whilst propionate decreased with some fluctuation. Never-

theless, total VFA concentrations in R1 remained at low levels.

In R2, a sharp decrease in acetate was noticed, the concentra-

tions of propionate and butyrate however increased during

the initial 12 and 5 days, respectively. On the contrary, acetate

concentration in R3 remained much higher than the values of

propionate and butyrate. As a simultaneous response to

inhibition, the butyrate concentration in R3 was observed to

increase from the initial 0.17 to 4.9 g/L within the first 7 d.

Thereafter, the concentration decreased with time. The

acetate concentration on the other hand fluctuated from 2.4

to 3.6 g/L during the first 5 d and later decreased promptly to

very low levels within 2 days. The increase in propionate

concentration was found to be around day 5, which increased

sharply and reached 1.0 g/L on day 12 but later decreased

slowly with time. Thus in this study, the butyrate rather than

the propionate or acetate, was found to be the most

important parameter for process monitoring. These results

were not in accord with previous studies, where the first sign

of reactor overload was reported to be either due to a

chronical build-up of propionic acid in the CSTR process

ARTICLE IN PRESS

WAT E R R E S E A R C H 4 0 ( 2 0 0 6 ) 2 6 2 1 – 2 6 2 82626

(Marchaim and Krause, 1993), or acetic acid accumulation in

both CSTRs and reactors with attached growth (Bolzonella et

al., 2003b; Cobb and Hill, 1991). Nevertheless, this observation

reaffirms the suggestion made by Ahring et al. (1995), that

VFA concentration during the start-up process is the best and

most reliable parameter to evaluate process stability.

The present results thus suggests that thermophilic CSTR

for treating SS-OFMSW and other high-strength substrates

can be started up successfully and relatively fast provided

thermophilic inoculum can be supplied and a careful loading

procedure is used, such as the ‘‘activated biomass’’ concept

used for R2 above. A similar start-up strategy such as use of

seed material from plants treating similar waste and gradual

organic load increase were reported (e.g., Bolzonella et al.,

2003a; Fernandez et al., 2001; Van Lier et al., 1993). Based on

the ‘‘activated biomass’’ concept, substrate could be added

progressively in a controlled manner until the designed

working volume was reached. Once the designed working

volume is attained, daily feeding and extraction can be

initiated and final design loading rate should be possible

shortly after. In practice (start-up of full-scale plants), the

‘‘activated biomass’’ concept can be adapted to handle also a

gradual supply of inoculum during the initial operating

period, as inoculum may only be available in limited daily

batches (normal daily extraction amount) from other plants

and without disturbing its operation. The controlled feeding

according to the ‘‘activated biomass’’ concept, designed to

avoid accumulation of intermediates, is also ideal to allow for

0

2

4

6

0 15 30 45

Time (d

TV

FA

(g/

L)

0

3

6

9

Bio

gasP

rod.

(100

0 m

3 /d

)

0.0

0.3

0.5

0.8

1.0

Sp.

bio

gas

Pro

d.(m

3 /k

gVS

)

Fig. 3 – Biogas production (J) and filling volume (’), specific bio

acids (TVFA: titration method; J reactor 1, ’ reactor 2 and K re

centralized biogas plant, Denmark) with daily loading at 5% of ca

concept’’.

less frequent VFA sampling/monitoring, as the loading

philosophy in itself should ensure against major imbalance

occurring between VFA analyses, while periodic VFA analyses

are sufficient to adjust the relative loading rate according to

actual process capacity.

3.4. Start-up of full-scale plant based on the ‘‘activatedbiomass’’ concept.

The start-up process performance of a thermophilic full-scale

plant is illustrated in Fig. 3. Reactor R1 (the first reactor to be

started-up) showed to respond well to the loading strategy

based on the feed activation concept. Following the batch-fed

mode filling up period (58 days), stable biogas production was

noticed with a pH around 8 and VFA content o3 g/L (based on

titration). The low VFA content during and after the start-up

period reaffirmed that the initial low-feeding strategy ap-

proach has avoided possible VFA build-up through a gradual

acclimation of the biomass. Filling the other two reactors

(R2 and R3) by splitting the R1 effluent did not affect the start-

up in either R2 or R3 and VFA content in the respective

reactors remained o4 g/L (Fig. 3). Throughout the start-up

period, pH remained in the region of 8 (70.2) and the

ammonia/ammonium concentration only increased slightly

from an initial level of approx. 3 g/L to a stable level of approx.

4 g/L at the end of the start-up period. These results indicate

that the inoculum, apart from being a thermophilic, had

also similar chemical/ionic properties ensuring a smooth

60 75 90

ays)

0

3

6

9

12

15

Fill

ing

Vol

(100

0 m

3 )

0.0

0.3

0.6

0.9

Load

ing

rate

(m3

/m3 )

gas production (J) and loading rate (–) and total volatile fatty

actor 3) during the start-up of full-scale biogas plant (Limvig

lculated active biomass according to the ‘‘activated biomass

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WAT E R R E S E A R C H 40 (2006) 2621– 2628 2627

adaptation to the fresh feed supplied to the new plant. This

example on the application of a controlled start-up loading

procedure for a full-scale thermophilic process, which has

been repeated with success on later plants, demonstrates that

process stabilization can be smooth allowing full attention to

be focused on other technical aspects of starting up a new

plant with the aim of reaching full design production

(income) as fast as possible.

4. Conclusion

Results from batch experiments showed that maximum

methane yields (0.43 m3/kgVSadded waste) and best degradation

rates for SS-OFMSW could be achieved at relatively low fresh

TS content (1.5%) and under thermophilic conditions. At

higher TS (3–30%), the process would be inhibited initially and

the magnitude of the inhibition would increase with increase

in TS.

The study also demonstrated that full-scale plants to treat

SS-OFHSW can be started-up successfully and relatively fast

provided at least 10–15% of final inoculum volume was

supplied and a careful loading procedure such as the

‘‘activated biomass’’ concept was adopted. Low and progres-

sively increasing loading (1.7–15 gVS/d) was found to be an

ideal start-up strategy compared to high constant loading rate

(5.7 gVS/d). Thus, a controlled start-up loading procedure

would not only allow rapid process stabilization but also

provide time to focus on other technical aspects of new plant

operation with the aim of reaching full design production

(income) as fast as possible.

Acknowledgement

Danish Research Council (FTP) is greatly acknowledged for

partial funding of this project (33031-0029/EFP-05).

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