Professional Documents
Culture Documents
Available at www.sciencedirect.com
http://www.elsevier.com/locate/biombioe
Article history: Cattle excreta and two-phase olive mill wastes (TPOMW) were codigested at a 3:1 ratio in
Received 28 April 2009 two 75 L continuous stirred tank reactors at 37 C and 55 C to analyse their biogas
Received in revised form production. The contribution of each residue to the total gas production at 37 C was
6 November 2009 evaluated in reactors digesting either 3:1 excreta:water or 3:1 water:TPOMW. The meso-
Accepted 11 November 2009 philic co-fermentation of cattle excreta with TPOMW at an organic loading rate (OLR) of
Available online 3 December 2009 5.5 g COD L1 d1 rendered 1096 mL biogas L1 sludge d1. This was 337% higher than that
of excreta alone. The methane yield resulting from the codigestion was 179 L CH4 kg1 VS
Keywords: loaded, of which 42% was attributed to the quarter of the reactor corresponding to TPOMW.
Animal waste Under thermophilic conditions, the codigestion yielded 17.3% more methane than meso-
Codigestion philically. In the reactor digesting TPOMW alone (OLR ¼ 3.8 g COD L1 d1) the ratio VFA/
Biogas alkalinity exceeded 0.8 after 21 d, leading to its acidification and inhibition of methano-
Methane genesis. Farm-scale digestion of animal excreta and TPOMW should be promoted in
Two-phase olive mill wastes Mediterranean countries as an environmentally sound option for waste recycling and
renewable energy production.
ª 2009 Elsevier Ltd. All rights reserved.
* Corresponding author. Tel.: þ43 512 507 5995; fax: þ43 512 507 2928.
E-mail address: marta.goberna@uibk.ac.at (M. Goberna).
0961-9534/$ – see front matter ª 2009 Elsevier Ltd. All rights reserved.
doi:10.1016/j.biombioe.2009.11.005
biomass and bioenergy 34 (2010) 340–346 341
EX_OL37 3:1 excreta:TPOMW 37 3.5 5.5 All reactors were run at 37 C and fed daily with cattle excreta
EX_OL55 3:1 excreta:TPOMW 55 3.5 5.5 at an equivalent feeding rate until a stable and similar biogas
EX37 3:1 excreta:water 37 3.5 1.4 production was reached, averaging 18.1 0.4 L d1
OL37 3:1 water:TPOMW 37 3.5 3.8 (mean SD) with a methane content of 68.7 0.4% v/v. This
a TPOMW: Two-phase Olive Mill Wastes. steady state was maintained for three days (days 1–3 in Fig. 2)
b FFR: Feed Flow Rate. prior to the start of the experiment. In day 3, the temperature
c OLR: Organic Loading Rate. of reactor EX_OL55 was set to 55 C. In day 4 (solid line in
Fig. 2), the feeding with specific solutions was initiated (Tables
1 and 2).
Immediately after starting the specific load, reactor
the mill processing effluent line. Both materials were stored at EX_OL37 showed an increase in biogas production (Fig. 2). This
4 C until reactor start up on 19 December 2007. stabilised at day 8, i.e. four days after starting the load with
specific feeding solutions, until the end of the experiment
averaging 82.0 4.8 L d1. Methane content in the biogas was
2.2. Sampling of gas and sludge constant, with 62.8 1.2%. In EX_OL55, there was an increase
in biogas production with lower methane content from days
Reactors were operated with the specific feed solutions for 3–4, which was a direct effect from the temperature rise to
30 d. The volume of biogas produced was quantified using 55 C (Fig. 2). This was followed by a decrease in biogas
a gas meter located on top of each reactor (Fig. 1). Gas samples production and its CH4 content from days 4–7, which
were collected daily in gas-tight bags and gas composition progressively recovered from day 8 up to day 14 when it sta-
(CH4, CO2 and H2) measured using a Biogas Monitor BM 2000 bilised at 94.5 10.5 L biogas d1 with 62.0 1.5% methane
(Geotechnical Instruments, Leamington, UK). Three sludge content. During the stable period, non-stirring conditions due
samples (ca. 500 ml) were collected every 3.5 d from a tap to technical problems occurred, which resulted in a slight
located in the bottom of each reactor (Fig. 1). decrease in biogas production (dashed lines in Fig. 2). Reactor
EX37 remained stable throughout the experiment, its biogas
production averaging 17.7 1.7 L d1 with 64.8 1.2% CH4
(Fig. 2). Finally, reactor OL37 rapidly reacted to the initial
2.3. Analytic methods change in feed, reaching a maximum biogas production at day
8 (Fig. 2). Biogas production remained relatively high from
Total solids (TS), volatile solids (VS), pH and electrical days 7–14 (59.5 3.1 L d1) and then decreased rapidly.
conductivity (EC) were determined following EU Horizontal Table 3 summarises the gas production parameters during
standards [20]. Total C and N contents were measured in an the steady state after the period corresponding to one HRT
automated CNHS analyser (TruSpec, LECO, Michigan, USA). (21.4 d). Data are average of three consecutive sampling dates.
Total and soluble chemical oxygen demand (COD) were ana- Reactor EX_OL37 produced 337% more biogas and 73.3%
lysed with the LCK 014 test kit and ammonia nitrogen (NH4–N) higher methane yield than EX37. Reactor EX_OL55 showed the
with the LCK 303 test kit (both from Hach Lange, Düsseldorf, highest biogas production and methane yield. These were,
Germany). Particulate COD was calculated as the difference respectively, 17.6% and 17.3% higher than those of EX_OL37.
between total and soluble COD. Determination of volatile fatty During this steady state, reactors EX_OL37 and EX_OL55 were
acids (VFAs) was performed within 24 h of sampling. Briefly, the most efficient in VS removal.
15 ml of sludge were centrifuged (14,000 rpm, 30 min) and the
0.2 mm filtered supernatant analysed for VFAs by High Pres-
3.2. Effluent sludge properties
sure Liquid Chromatography (HPLC Shimadzu LC-20A promi-
nence) using 20 ml injection volumes. VFAs were separated
At the start of the experiment, while feeding was performed
through an ion exclusion column (300 7.8 mm, Aminex
with excreta alone, the amount of total solids (TS) in the
HPX-87H) and photometrically detected at 210 nm. An iso-
effluent sludge of all reactors was approximately 4.0 0.1%, of
cratic flow (0.7 ml min1) with 5 mM H2SO4 was used as the
which 66.5 0.8% were volatile solids (VS). Total and soluble
running medium at 60 C for 90 min.
chemical oxygen demand (COD) averaged 27.0 0.6 g L1 and
7 0.7 g L1, respectively. TS, VS, total and soluble COD
increased together with the change in feed in the effluent
2.4. Multivariate analysis sludge of reactors EX_OL37, EX_OL55 and OL37 (except for TS
in OL37), whereas all parameters slightly decreased in EX37
Principal component analysis (PCA) based on a correlation (Table 3).
matrix was performed to inspect the relationship between The main volatile fatty acids (VFAs) detected in all reactors
biogas production, effluent sludge properties and the factors were acetate and propionate (Fig. 3). VFAs showed relatively
‘‘reactor’’ and ‘‘sampling date’’, using SPSS 15.0 for Windows. constant values in reactors EX_OL37 and EX37, their levels
biomass and bioenergy 34 (2010) 340–346 343
Fig. 2 – Biogas production (L dL1) and methane content in the biogas (% v/v) in the reactors (EX_OL37, 3:1 excreta:TPOMW at
37 8C; EX_OL55, 3:1 excreta:TPOMW at 55 8C; EX37, 3:1 excreta:water at 37 8C; OL37, 3:1 water:TPOMW at 37 8C). Solid line
indicates start of feeding with specific mixtures. Dashed lines indicate period under non-stirring conditions.
being always below 0.8 and 0.2 g VFA L1, respectively. Both the experiment to values averaging 0.9 0.03 g L1 and
reactors showed stable pH values above 7.6 throughout the 1.1 0.05 g L1, respectively. In EX37, NH4–N oscillated
experiment (Fig. 3). However, EX_OL55 and OL 37 reached between 1.1 and 1.3 g L1, whereas in OL37 it decreased
peaks of 4.8 and 3.8 g VFA L1. The increase in effluent VFAs in progressively to final values of 0.5 g L1. Alkalinity was highly
EX_OL55 was mainly due to the accumulation of acetate, and correlated with NH4–N (Pearson’s correlation
to a lesser extent propionate, after temperature was changed coefficient ¼ 0.80, p < 0.001) and followed similar temporal
from 37 C to 55 C on day 3. A subsequent decrease in pH was patterns (Fig. 4).
detected from an initial 7.8 down to 7.4. This reverted on day
13 after acetate levels started to stabilise. In reactor OL37, both 3.3. Principal component analysis
acetate and propionate progressively accumulated, which
was accompanied by a decrease in pH down to 6.5 (Fig. 3). The two principal components (PCs) extracted from the ordi-
Total ammonia nitrogen (NH4–N) averaged 1.3 0.03 g L1 nation of the data explained 76.6% of the total variance. Fig. 5A
in all reactors before the shift to the specific feed solutions on shows the distribution of the samples within the biplot that
day 4 (Fig. 4). In EX_OL37 and EX_OL55, NH4–N decreased on both PCs define. At the start of the experiment (day 4), all
day 11 (i.e. five days after changing the feed) until the end of reactors were located near to each other in the negative pole of
PC1, which was highly correlated with ammonia N (Fig. 5B). As
the digestion with the specific feed solutions proceeded, the
Table 3 – Steady state parameters after the period reactors followed divergent trajectories in the diagram. Both
corresponding to one HRT (21.4 d). Standard deviations reactors codigesting excreta and TPOMW (EX_OL37 and
are given in brackets for n [ 3. EX_OL55) followed similar trajectories irrespective of their
Parametera EX_OL37 EX_OL55 EX37 OL37 operating temperature. These moved towards the area of high
biogas production, and high levels of TS and COD, responding to
Gas production
the change of feed from excreta alone (OLR ¼ 1.9 g COD L1 d1)
Biogas production 1096 (81) 1289 (158) 251 (39) –
to excreta:TPOMW (OLR ¼ 5.5 g COD L1 d1). EX37 showed the
(mL L1 sludge d1)
Methane yield (L CH4 kg1 179 (18) 210 (30) 103 (16) – shortest trajectory, thus the most similar performance
VS added) throughout the experiment, towards the sector of low TS and
VS removed (%) 53.4 (1.7) 54.0 (1.8) 45.8 (6.3) – COD, due to the decrease in OLR from 1.9 (excreta) to
Effluent characteristics 1.4 g COD L1 d1 (diluted excreta). Finally, OL37 advanced in the
TS (% w/v) 5.2 (0.1) 5.2 (0.2) 2.9 (0.2) 3.4 (0.1) direction of the area correlated with low biogas production,
VS (% w/w of TS) 73.6 (0.6) 73.4 (0.6) 62.3 (1.9) 72.6 (1.5) alkalinity, pH and ammonia N, but high total C, total N and
Particulate COD (g L1) 27.9 (0.2) 26.0 (0.6) 16.5 (0.5) 22.4 (1.1) propionate.
Soluble COD (g L1) 10.9 (0.7) 16.9 (0.9) 5.5 (0.2) 13.1 (0.7)
Total C (g L1) 41.5 (2.4) 44.1 (3.6) 41.7 (2.2) 46.4 (2.5)
Total N (g L1) 3.5 (0.1) 3.5 (0.2) 3.2 (0.3) 3.4 (0.2)
EC (mS cm1) 16.1 (0.1) 16.7 (0.3) 15.3 (0.2) 10.8 (1.4) 4. Discussion
a TS, total solids; VS, volatile solids; COD, chemical oxygen
Four pilot reactors were used to evaluate the potential biogas
demand; EC, electrical conductivity.
production from the codigestion of cattle excreta and two-
344 biomass and bioenergy 34 (2010) 340–346
Fig. 3 – Acetate, propionate (g LL1) and pH in the reactors (EX_OL37, 3:1 excreta:TPOMW at 37 8C; EX_OL55, 3:1
excreta:TPOMW at 55 8C; EX37, 3:1 excreta:water at 37 8C; OL37, 3:1 water:TPOMW at 37 8C).
phase olive mill wastes (TPOMW). Prior to the start of the filled with TPOMW instead of water. These values are well
experiment with specific feeding solutions and temperatures, above those reported previously in experiments using olive
the reactors were run with cattle excreta at 37 C until they mill wastewaters. Gelegenis et al. [14] obtained
operated similarly in terms of gas production, methane 520 mL biogas L1 reactor d1 from the digestion of poultry
content and sludge properties (day 4 in Fig. 5A). This demon- excreta and olive mill wastewasters (pH 1.6) at a ratio 3:1 in
strated that all four reactors function similarly if operated a 25 L reactor. This value was 25% higher than the biogas
under equal conditions. The diverse combinations of wastes produced by a similar reactor fed only with poultry excreta. Up
led to distinct reactor performance (Fig. 5). to 155% additional biogas was obtained in a batch experiment
The reactors codigesting excreta and TPOMW at 37 C in 100 ml vials when laying hen litter was digested with olive
(EX_OL37) and 55 C (EX_OL55), and the reactor digesting mill effluent compared to laying hen litter alone, at
excreta alone (EX37) operated stably and achieved a constant 10 g COD L1 load with 10% TS [16].
biogas production and methane yield. After the period corre- The methane yield of the mesophilic reactor codigesting
sponding to one HRT (21.4 d), 1096 mL L1 d1 biogas were cattle excreta and TPOMW at a ratio 3:1 was 179 L CH4 kg1 VS
produced from the sludge in EX_OL37 while EX37 yielded only loaded. A total 57.7% of this methane can be attributed to the
251 mL L1 d1. Therefore, 337% more biogas was produced three quarters of the reactor that were filled up with excreta,
when a quarter of the effective volume of the digester was considering the results from the excreta digesting reactor
Fig. 4 – Ammonia N (mg LL1) and alkalinity (mmol HD equiv LL1) in the reactors (EX_OL37, 3:1 excreta:TPOMW at 37 8C;
EX_OL55, 3:1 excreta:TPOMW at 55 8C; EX37, 3:1 excreta:water at 37 8C; OL37, 3:1 water:TPOMW at 37 8C).
biomass and bioenergy 34 (2010) 340–346 345
cattle excreta:two-phase olive mill wastes with an OLR of [10] Beccari M, Majone M, Riccardi C, Savarese F, Torrisi L.
5.5 g COD L1 d1 and a HRT of 21.4 d. The quarter of the Integrated treatment of olive oil mill effluents: effect of
reactor filled up with TPOMW, with its high biodegradability, chemical and physical pretreatment on anaerobic
treatability. Water Sci Technol 1999;40:347–55.
contributed to 75.6 L CH4 kg1 VS of the total methane yield.
[11] Hamdi M. Anaerobic digestion of olive mill wastewaters.
The excreta buffered the low pH and alkalinity of the TPOMW, Process Biochem 1996;31:105–10.
allowing its anaerobic biodegradation without the need of any [12] Borja R, Rincón B, Raposo F, Alba J, Martı́n A. A study of
pre-treatment or chemical additive. Thermophilically, the anaerobic digestibility of two-phases olive mill solid waste
codigestion of both wastes rendered 17.3% more methane (OMSW) at mesophilic temperature. Process Biochem 2002;
than mesophilically. Reactor efficiency in terms of VS removal 38:733–42.
[13] Ergüder TH, Güven E, Demirer GN. Anaerobic treatment of
was higher (ca. 54%) thanks to the codigestion of the residues.
olive mill wastes in batch reactors. Process Biochem 2000;36:
The mixing ration should be optimized according to the
243–8.
precise composition of the input materials. [14] Gelegenis J, Georgakakis D, Angelidaki I, Christopoulou N,
Goumenaki M. Optimization of biogas production from olive-
oil mill wastewater, by codigesting with diluted poultry-
Acknowledgements manure. Appl Energ 2007;84:646–63.
[15] Fountoulakis MS, Drakopoulou S, Terzakis S, Georgaki E,
We thank the Agricultural School in Rotholz (Tirol, Austria) Manios T. Potential for methane production from typical
Mediterranean agro-industrial by-products. Biomass
and the cooperative COATO in Totana (Murcia, Spain) for
Bioenerg 2008;32:155–61.
providing us with the cattle excreta and TPOMW, respectively. [16] Azbar N, Keskin T, Yuruyen A. Enhancement of biogas
We also thank M.A. Sánchez Monedero, A. Roig and C. Mon- production from olive mill effluent (OME) by co-digestion.
dini for arranging the transport of TPOMW to our laboratory. Biomass Bioenerg 2008;32:1195–201.
We thank C. Mondini for providing nutrient and heavy metal [17] Schoen MA, Sperl D, Gadermaier M, Goberna M, Franke-
contents of the digestion end products and E.A. Eladawy for Whittle I, Insam H, Wett B. Comparison of biogas plant start-
providing Fig. 1. M. Goberna was supported by the Marie Curie up procedures based on lab- and full-scale data and on
numerical modelling. In: Proceedings of the Second
Actions (MEIF-CT-2006-041034). The support of the Tiroler
International Symposium on Energy from Biomass and
Zukunftsstiftung for the K-Regio Center BioTreaT is Waste. Venice; 2008.
appreciated. [18] Schoen MA, Sperl D, Gadermaier M, Goberna M, Franke-
Whittle I, Insam H, et al. Population dynamics at
digester overload conditions. Bioresour Technol 2009;
references 100:5648–55.
[19] Wett B, Schoen MA, Phothilangka P, Wackerle F, Insam H.
Model-based design of an agricultural biogas plant:
[1] Tabajdi CS. Draft report on sustainable agriculture and application of anaerobic digestion model no.1 for an
biogas: a need for review of EU-legislation (2007/2107 INI). improved four chamber scheme. Water Sci Technol 2007;55:
Brussels: Committee on Agriculture and Rural Development, 21–8.
European Parliament; 2007. [20] Van der Sloot H (coord.) Project horizontal: European
[2] US Environmental Protection Agency. Anaerobic digestion: standards in the field of soil, sludge and treated biowaste on
benefits for waste management, agriculture, energy, and the sampling, inorganic parameters, organic parameters,
environment. US Environmental Protection Agency, http:// biological parameters and hygienic parameters, CEN/BT Task
www.dcmnr.gov.ie/NR/rdonlyres/287C17F6-13D2-48B9- Force 51. ULR: http://www.ecn.nl/horizontal/; 2006.
882C-2060512A573E/0/EPAappendix.pdf; 2005. [21] Güngör-Demirci G, Demirer GN. Effect of initial COD
[3] European Parliament. Sustainable agriculture and biogas: concentration, nutrient addition, temperature and microbial
a need for review of EU-legislation. Non-legislative resolution acclimation on anaerobic treatability of broiler and cattle
T6-0095/2008. Brussels: European Parliament; 2008. excreta. Bioresour Technol 2004;93:109–17.
[4] Angelidaki I, Ahring BK. Methods for increasing the biogas [22] Sánchez E, Borja R, Travieso L, Martin A, Colmenarejo MF.
potential from the recalcitrant organic matter contained in Effect of organic loading rate on the stability, operational
manure. Water Sci Technol 2000;41:189–94. parameters and performance of a secondary upflow
[5] Demirer GN, Chen S. Anaerobic digestion of dairy manure in anaerobic sludge bed reactor treating piggery waste.
a hybrid reactor with biogas recirculation. World J Microb Bioresour Technol 2005;96:335–44.
Biot 2005;21:1509–14. [23] Zhao HW, Viraraghavan T. Analysis of the performance of an
[6] Bayerische Landesanstalt für Landwirtschaft. anaerobic digestion system at the Regina wastewater
Biogasausbeuten verschiedener substrate. Bayerische treatment plant. Bioresour Technol 2004;95:301–7.
Landesanstalt für Landwirtschaft, http://www.lfl.bayern.de/ [24] Mondini C. Nutrient and heavy metal content of effluent
ilb/technik/10225/; 2004. sludge from reactors digesting cattle excreta and two-phase
[7] Borja R, Rincón B, Raposo F. Anaerobic biodegradation of olive mill waste. Gorizia, Italy: Centro di ricerca per lo studio
two-phase olive mill solid wastes and liquid effluents: kinetic delle relazioni trapiantae suolo, pers.com; April 2009.
studies and process performance. J Chem Technol Biot 2006; [25] Karri S, Sierra-Alvarez R, Field JA. Toxicity of copper to
81:1450–62. acetoclastic and hydrogenotrophic activities of methanogens
[8] Roig A, Cayuela ML, Sánchez-Monedero MA. An overview on and sulfate reducers in anaerobic sludge. Chemosphere 2006;
olive mill wastes and their valorisation methods. Waste 62:121–7.
Manage 2006;26:960–9. [26] Eladawy EA. Modelling of anaerobic sewage sludge digestion –
[9] Angelidaki I, Ahring BK. Codigestion of olive oil mill Sludge characterisation and process analysis. Innsbruck:
wastewaters with manure, household waste or sewage Institute of Environmental Engineering, University of
sludge. Biodegradation 1997;8:221–6. Innsbruck. PhD thesis; 2005.