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Bioresource Technology 101 (2010) 545550

Contents lists available at ScienceDirect

Bioresource Technology
journal homepage: www.elsevier.com/locate/biortech

Case Study

Thermophilic anaerobic co-digestion of cattle manure with agro-wastes

and energy crops: Comparison of pilot and full scale experiences
C. Cavinato a,*, F. Fatone b, D. Bolzonella c, P. Pavan a
a

Department of Environmental Sciences, University Ca Foscari of Venice, Dorsoduro 2137, I-30123 Venice, Italy
Department of Biotechnology, University of Verona, Strada Le Grazie 15, I-37134 Verona, Italy
c
Department of Science, Technology and Market of Wine, University of Verona, via della Pieve 70, 37020 San Floriano, Verona, Italy
b

a r t i c l e

i n f o

Article history:
Received 27 May 2009
Received in revised form 24 July 2009
Accepted 7 August 2009
Available online 10 September 2009
Keywords:
Anaerobic co-digestion
Thermophilic
Biogas
Cattle manure
Economics

a b s t r a c t
The paper deals with the benets coming from the application of a proper process temperature (55 C)
instead of a reduced thermophilic range (47 C), that is often applied in European anaerobic co-digestion
plants. The experimental work has pointed out that biogas production improve from 0.45 to 0.62
m3/kg VS operating at proper thermophilic conditions. Moreover, also methane content was higher: from
52% to 61%. A general improvement in digester behaviour was clear also considering the stability parameters comparison (pH, ammonia, VFA content). The second part of the study takes into account the
economic aspects related to the capital cost of anaerobic digestion treatment with a 1 MW co-generation
unit fro heat and power production (CHP). Moreover, the economic balance was also carried out considering the anaerobic supernatants treatment for nitrogen removal. The simulation showed how a payback-time of 2.5 yr and between 3 and 5 yr respectively could be determined when the two options of
anaerobic digestion only and together with the application of a nitrogen removal process were
considered.
2009 Elsevier Ltd. All rights reserved.

1. Introduction
The need to reduce the emissions of green house gases, especially carbon dioxide, and to develop a reliable alternative to the
fossil fuel economy, is conveying the interest of policy makers toward the renewable energy sources. In fact, together with the increased efciency in the energy sector (that is a reduction of
consumption), renewable energy sources can contribute to the
reduction in fossil fuel using and carbon dioxide emissions. Beside
solar, hydro, wind or geothermal-energy, the biogas production
from organic waste is knowing a renaissance after the interest of
the past (Ghosh and Pohland, 1974; Cecchi and Traverso, 1988;
Chynoweth et al., 1990; Kayhanian and Tchobanoglous, 1993; Cecchi et al., 1994). Anaerobic co-digestion of agricultural wastes and
energy crops, in particular, is supposed to be one of the main alternative in this sector, as stated also by the United Nations Development Programme (UNPD) that consider this technology as one of
the most useful decentralised sources of energy supply, especially
if used with energy crops and all substrates easily available in
many farms. Moreover, considering the complete waste-to-energy
transformation, anaerobic processes can be considered a way to reduce the organic content of biowaste giving low-CO2 emission.

* Corresponding author. Tel.: +39 0422 321037; fax: +39 0422 326498.
E-mail address: cavinato@unive.it (C. Cavinato).
0960-8524/$ - see front matter 2009 Elsevier Ltd. All rights reserved.
doi:10.1016/j.biortech.2009.08.043

Agricultural wastes can be considered as a primary substrate for

these aims, in particular considering co-digestion of manure and
other specic biomass coming from cultivations. Manure, in particular, is a resource easily available in many farms all over the world.
However, the reduced biogas yield of this material, sometimes
does not justify the capital costs for farm-scale plants. However,
the biogas productivity can be dramatically increased by introducing energy rich co-substrates to the anaerobic digester (maize,
grass, bread, fruit, etc.).
Several experiences have shown how the thermophilic range of
temperature should be preferred for the co-digestion process because of its superior performances compared to a mesophilic process (Mladenovska and Ahring, 2000; Ahring et al., 2001; Van Lier
et al., 2001; Angelidaki et al., 2006) as well as its sanitisation
capability.
In central and north Europe, in particular, anaerobic digestion is
widely applied in the agricultural sector. In Denmark, for example,
there are some 20 centralised biogas plants and more than 60
farm-scale plants treating livestock manure. Most of the centralised biogas plants treat manure together with other organic
wastes; the preferred temperature range is thermophilic and the
HRT is 1122 d (Nielsen and Angelidaki, 2008). The annual amount
of manure and other biomass treated is about 1.5 million ton/yr,
producing a biogas equivalent of about 39,000,000 m3 CH4/yr
(Angelidaki and Ellegaard, 2003).

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C. Cavinato et al. / Bioresource Technology 101 (2010) 545550

Also in Sweden the choice of co-digestion was implemented in

more than 200 sites: ten of them are centralised plants where
manure is co-digested together with various kind of waste, usually
originated from the food-processing industries or the source separated collection of restaurant wastes (Lantz et al., 2007), all the
others are farm-scale plants treating manure and crop residues.
This situation can be ascribed to the fact that Sweden is so spread
out that full utilisation of this energy potential by centralised slurry based technology is difcult (Svensson et al., 2005), therefore,
decentralised plants are preferred.
Usually, on farms, bioreactors may be subjected to temperature
uctuations due to large variations in outdoor temperature and
reactor feeding, especially in highland and northern climates, or
non optimal set-up ranges. All these problems can cause instability
and disturbances in all the main parameters of the process (Alvarez
and Lid, 2008; Lettinga, 2004; Mass et al., 2003), leading to notable yields reduction.
As a general remark we can say that often a not optimal temperature range is applied in these plants.
Ahring (1994) showed that the optimal temperature for thermophilic digestion was found to be 60 C. However, for practical
operation of full scale plants, a temperature between 52 and
56 C will be preferable, allowing a slight variation in the temperature without fatal consequences for some of the active microbes.
Following this evidences, this paper consider the anaerobic codigestion of manure with other agro-waste in mesophilic and thermophilic conditions at both pilot and full scale: in particular the results coming from a 0.38 m3 stirred reactor operating at 55 C are
compared to those coming from a 1400 m3, two-stage process,
operating at 47 C.
2. Methods
The experimental plan was divided in two phases. During the
rst part, the operational conditions of a full scale plant were
reproduced at pilot scale in order to study the stability of the process under the same temperature conditions (47 C). On the other
hand, during the second part of the experimental work, the temperature of the pilot-scale process was increased at 55 C, with
the aim of improving performances and process yields.

2.1. Full scale plant conguration

The full scale plant based in Marcon-Venice and shown in Fig. 1,
was composed by a mechanical feed system for manure, maize and
other food waste residues (bread, fruit, grass, etc.), a storage tank
(900 m3 volume) for the liquid manure feeding the reactor and also
for the recycle operations, an anaerobic digester with volume of
1400 m3 operating at an average temperature of 47 C, a storage
tank for treated manure (4300 m3 volume), a dewatering system
and an unit for the co-generation of heat and power (340 kW h
CHP unit).
The plant treated 140,000 kg/d of cattle manure mixed with
agro-industrial wastes and 25 m3/d of liquid manure (inclusive of
efuent recycling). The organic loading rate (OLR) applied was
some 5 kg TVS/m3 d with a HRT of some 34 d while the storage
tank allowed for other 3 months of retention time for further
stabilisation.
2.2. The pilot-scale plant
The experiment was carried out in a CSTR reactor of stainless
steel, with a working volume of 380 l. The reactor was heated by
hot water circulation. The reactor was inoculated with the anaerobic sludge coming from the full scale plant of Marcon (122 g/kg TS
and 92 g/kg TVS) in order to reproduce the same conditions of the
full scale reactor.
2.3. Analytical schedule and methods
The efuent of the full scale plant was monitored 2 times/week
while the pilot reactor was monitored 5 times/week.
The efuent of both reactors was monitored in terms of total
(TS) and volatile solids (VS) content, chemical oxygen demand
(COD), total Kjiendhal nitrogen (TKN), total phosphorus (TP), and
stability parameters (pH, alkalinity, ammonia and volatile fatty
acids (VFA) content), all in accordance with the Standard Methods.
The VFA content was monitored using a gas chromatograph
(Carlo Erba instruments) with hydrogen as gas carrier, equipped
with a Fused Silica Capillary Column (Supelco NUKOLTM,
15 m  0.53 mm  0.5 lm lm thickness) and with a ame ioniza-

Fig. 1. Simplied ow diagram of the full scale plant.

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C. Cavinato et al. / Bioresource Technology 101 (2010) 545550

Table 1
Characterisation of inuent substrates.
Substrate

TS, g TS/kg w.w.

TVS, g TVS/kg w.w.

VS, %

COD, g COD/kg TS

TKN, g N/kg TS

Ptot, g PPO4/kg TS

VFA, mg COD/l

NNH3, mg N/l

Solid manure
Liquid manure
Maize
Fruit
Bread

361
97
351
219
828

304
74
327
211
748

84
76
93
96
90

904
877
1061
1091
1143

30.3
34.2
16.4
16.6
24.8

4.33
12.00
2.35
1.88
1.06

25.9
756.6
40.5
910.6
n.d.

4482

56
55

6
5

53

52
51

50

49

54
kgTVS/m3d

tion detector (200 C). The temperature during the analysis started
from 80 C and reach 200 C trough two other steps at 140 and
160 C, with a rate of 10 C/min. The analyzed samples were centrifuged and ltrated with a 0.45 lm membrane. Gas production in
the pilot plant was monitored continuously and on line by a gas
ow meter (Ritter Company, drum-type wet-test volumetric gas
meters), while the biogas composition (CO2CH4H2S) was dened
by a portable infrared gas analyser (geotechnical instrument,
model GA2000).

48
1
OLR

2.4. Substrates characterisation

temperature

The reactors were fed with a mixture of cattle manure (solid

and liquid), maize, fruit-processing waste (marc) and bread of
27%, 18%, 37% and 18%, respectively, on wet weight basis, to obtain
a suitable solid content in the digester feeding (some 1012% total
solids).
Table 1 shows the average values found for the characterisation
of each substrate.
With reference to data shown in Table 1, the solid fraction of
manure and maize showed similar characteristics, with a TS content of some 3536% and a volatile fraction of 84% and 93%, respectively, while fruit showed a lower content in terms of total solids
(22%) but a higher VS concentration (96%). As for bread, this
showed a solid content of some 83%, 90% volatile while the liquid
fraction of manure showed a TS content <10%. All the substrates
were characterized by a COD/TS ratio in the range 0.91.0. As for
the nutrients content it turned out clearly that manure was
responsible for high nitrogen and phosphorus contents. Liquid
manure, in particular, showed high presence of ammonianitrogen: some 45 g/l.
3. Results and discussion
3.1. Anaerobic co-digestion at pilot and full scale
As mentioned above, the pilot scale test was divided in two
periods, the rst carried out at 47 C and the second at 55 C. For
both those temperatures, were reached stable steady state conditions, however the high variability of the substrates characteristics
inuenced the stability of the process (Fig. 2).
The organic loading rate applied to the pilot scale reactor was
maintained at about 5 kg TVS/m3 d (as in the full scale) during
the steady state periods.
As for temperature changes, the most common strategies of
adapting mesophilic reactors to thermophilic temperatures are
two: a one-step or a step-wise temperature increase. These were
both widely described in literature and were often applied in real
cases (Bouskova et al., 2005). Nevertheless, there is a lack of studies
where strategies for temperature change are compared (from mesophilic to thermophilic range), especially treating maize, cattle
manure and other similar organic substrates.
In this experimental work, according to Cecchi et al. (1993), a
quick change of thermal conditions was performed and the digester feeding interrupted in order to not upset the bacterial food-

47
46

20

40

60

80

100

120

140

days
Fig. 2. OLR behaviour during the experimental work, with a focus on the change
temperature period.

Table 2
Characteristics of digested sludge, operational conditions and process yield.
Full scale

Pilot plant

47 C

Run 1 (47 C)

Run 2 (55 C)

Characterisation of digested sludge

pH

7.94
mg N/l
4762
NNH3
VFA
mg COD/l
592
TS
g TS/kg w.w.
122.0
TVS
g TVS/kg w.w.
91.9

8.06
3647
1777
82.5
62.8

8.17
2429
313
83.4
64.1

Operational conditions
OLR
kg VS/m3 d
HRT
d

5.39
33.0

5.67
34.5

4.66
34.2

Process yield
GPR
m3/m3 d
SGP
m3/kg VS
%
CH4
ppm
H2S

2.4
0.45
52.3
884

2.9
0.54
58.8
483

2.7
0.62
61.6
549

chain (Bolzonella et al., 2003a,b). Fig. 2 shows this particular

situation.
Table 2 reports the main operational conditions and the process
yield as well as a detailed characterisation of the digested sludge.
Considering the period at 47 C and comparing the results at full
and pilot scale, it turns out clear that the two reactors operated at
similar OLR and HRT and the stability parameters were similar: in
particular, the pH showed an average value of eight and the same
was for ammonia concentration (some 4 g/l).
On the other hand, the TS and TVS content at pilot scale was
lower than the full scale while the VFA concentration was higher:
this let suppose a better degradation of the substrates, conrmed
also by the results in terms of biogas production.
In fact, the specic gas production was a slightly higher in the
pilot scale reactor than at full scale, 0.54 m3/kg TVSfed instead of
0.45 m3/kg TVSfed and with a methane content of 58.8% instead
of 52.3%. These better results can be probably ascribed to the good
mixing achieved in the pilot scale reactor: in fact, as already shown

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C. Cavinato et al. / Bioresource Technology 101 (2010) 545550

a 6000

SGP pilot plant

5000

m3/kgVS

mgN/l

4000
3000
2000
1000

N-NH4 digestor

N-NH4 liquid manure

0
0

20

40

60

80

100

120

140

20

40

60

days

b 8.5

SGP full scale

0.7
0.7
0.6
0.6
0.5
0.5
0.4
0.4
0.3
0.3
0.2

% CH4 pilot plant

80
days

100

120

140

160

% CH4 full scale plant

75
70

65
7.5

pH

60
7

55
50

6.5

pH digestor

pH liquid manure

45
40

6
0

20

40

60

80

100

120

140

days

35
0

Fig. 3. Ammonia concentration (a) and pH (b) of digested sludge and inuent liquid
manure in pilot plant.

by Stroot et al. (2000), this allowed for an improvement in the

substrates degradation and release of the biogas from the bulk,
resulting in an overall increased biogas production.
Ammonia and pH values of the pilot scale reactor were strongly
dependent on the substrates characteristics, in particular liquid
manure (Fig. 3a and b).
As for nitrogen concentration, which reached levels of some
45 g/l, it should be pointed out that at these pH and temperature
levels, the values for free ammonia (as NH3) were calculated in
0.84, 0.80 and 0.90 g N/l, respectively, for the full and pilot-scale
runs. These values were lower than the value found to cause inhibition by Hansen et al. (1998) and equal to 1.1 g N/l. On the other
hand, the authors themselves, experienced the operation of full
scale reactor operating at nitrogen concentrations of some 8 g N/l
in the bulk and producing some 180 m3 biogas per ton of raw biowaste treated, therefore showing that no inhibition was occurring
(Bolzonella et al., 2006).
From day 70 to day 110 (Fig. 4) the pilot plant specic gas production was variable. This behaviour was probably due both to the
strategy adopted to achieve the real thermophilic condition, and to
the great heterogeneity of inuent wastes combined to the feeding
strategy (i.e., whey with low TVS content, melon and water-melon
during summer period).
In the second pilot-scale run, carried out at 55 C, pH was constant (8.2) while ammonia concentration rstly decreased, mainly
because of the decrease of the inlet ammonia content, achieving an
average value of 2429 mg N/l. During the steady state conditions,
the specic gas production reached a SGP value of 0.62 m3/kg TVS
with a methane content of 61.6% despite a decrease in the organic
loading rate, which passed from some 5.5 to 4.7 kg TVS/m3 d. The
trends of the stability parameters and the yields obtained at
55 C, conrmed a better digester behaviour operating in these
conditions compared to the results obtained working at 47 C:
VFA decreased to some 300 mg/l compared to the mesophilic condition while TS and TVS concentrations in the reactor were the
same observed during the rst run.

20

40

60

80
days

100

120

140

Fig. 4. Comparison between biogas specic production (a) and methane content (b)
of pilot plant and full scale plant.

According to the results of this study it was shown that the

anaerobic co-digestion process carried out at 55 C showed an
increase of 15% in biogas production (SGP from 0.54 to 0.62
m3/kg TVS) while all the stability parameters (pH, ammonia and
VFA concentrations) showed the reliability of the process. Moreover, also the importance of a proper mixing (at pilot scale) was
emphasized.
3.2. Economics of the anaerobic co-digestion strategy
After the experimental evaluation of the optimum working
temperature, the economic aspects of the anaerobic co-digestion
strategy were considered both in terms of capital costs and of
efuent treatment costs and the net present value (NPV) was
determined.
To calculate the net present value was applied the following
equation:

NPVn C 0 bn  cn

1:035n  1
1:035n  0:035

where n is referred to the year considered, bn is the total amount of

annual benet, cn the total annual costs and C0 the capital cost.
In this paper the bn was determined as the benet coming from
the electric energy selling, also considering the credits for renewable energy (green certicates).
Taking into consideration the design parameters of the full scale
plant of Marcon and literature data (Bonazzi, 2001), the net present
value for implementation of the anaerobic digestion process in a
medium size farm was evaluated.
The average inlet ow treated was of 38 m3/d of liquid manure
from pigs/milk cows farming, and about 70 ton/d of maize. With
this loading conditions, and considering the optimal specic gas
production (at 55 C, 0.62 m3/kg TVS) the anaerobic process led
to a daily biogas production of 10,200 m3/d. After the evaluation
of the benet of the electric energy selling and the cost of the

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C. Cavinato et al. / Bioresource Technology 101 (2010) 545550

Table 3
Evaluation of capital cost and prot of a medium size biogas plant.

30%

Capital cost
m3 biogas produced
Cost EE + GC
Electric energy produced

m3/d
/kW h
MW h/yr

3,000,000
10,200
0.22
8789

Benet
Electric energy selling + benet from GC

/yr

1,933,473

Total

/yr

1,933,473

Staff (2 qualied labor)

Maize cultivation
EE used
General

/kW h
/yr
/yr
/yr
/yr
/yr

0.01
87,885
70,000
420,000
52,000
90,000

Total

/yr

719,885

Cost
Service CHP unit

60%
90%

9 10 11 12 13

years
Fig. 6. Net present value of the investment for the biogas plant considering the
efuent treatment cost for nitrogen removal.

In Fig. 6 are shown the net present values of the investment in

three different situations: removal of 30%, 60% or 90% of nitrogen
load in the anaerobic digestion efuent.
The same calculation of the net present value was made taking
into account the nitrogen removal cost. The pay-back time in the
rst condition (30% nitrogen removal) was similar to the initial
condition (3 yr); when the percentage of removed nitrogen rise
up to 90% the pay-back time became about twice the amount.
4. Conclusions

9 10 11 12 13

years
Fig. 5. Net present value of the investment for the biogas plant.

Green Certicates (0.22/kW h, Italian nancial law 2008), in Table

3 are reported the economics input and output for the estimation
of the net present value (Fig. 5).
The capital cost estimated was 3 million , inclusive of all the
equipments (among the others, 1 MW CHP unit and two digesters
with a total volume of about 5000 m3), but, considering the benet
coming from the electric energy selling, the net present value was
2.5 yr.
Following the Nitrates Directive (Commission of EC, 1991)
where vulnerability of the water bodies was taken into account,
the efuent treatment for nitrogen removal become the main
problem for farmers. It is well known that the anaerobic digestion
process convert the proteins organic nitrogen into high amount of
ammonia, and for cattle manure and similar waste, the average
ammonia content is about 44.5 g N/l (Hansen et al., 1998; Ahring
et al., 2001).
There are several papers in scientic literature where different
biological and chemical alternatives for nitrogen removal were
studied and compared, for example some authors (van Loosdrecht
and Salem, 2006; STOWA, 1996) have analysed the biological treatments of sludge digester liquids, comparing processes like the
SHARON process, ANNAMOX process, N-removal over nitrite, also
in economics terms. Bortone (2008) studied the N-removal efciency from piggery wastewater, through the integrated anaerobic/aerobic treatment showing the economics advantage of
coupling anaerobic digestion and SBRs.
These studies presented an average cost for nitrogen removal of
4/kg N removed: this value was used in this work to estimate the
annual cost for the digester efuent treatment. Considering the
same biogas plant data, the annual nitrogen production was
158 ton/yr.

The results of the present study showed that biogas production

from the co-digestion of cattle manure and other organic wastes
was increased when operating at proper thermophilic conditions
(55 C) and also a general improvement in digester behaviour is
clear considering the stability parameters. Further, the economic
aspects of the co-digestion strategy were considered: the results
showed that the net present value of the investment, considering
only the anaerobic digestion, was 2.5 yr. If the efuent treatment
for nitrogen removal was also considered in the calculation for a
30%, 60% and 90% efciency, the net present value of the investment observed was 35 yr, respectively.
Acknowledgements
This research was realized thanks to the funding of the Italian
Ministry of University and Research under the Project PRIN2007.
Andrettas farm, Marcon-Venice, where the full scale plant is located, is also acknowledged for the kind hospitality during the
experimental work.
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