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Anaerobic co-digestion of aquatic flora and quinoa with manures

from Bolivian Altiplano
René Alvarez a,*, Gunnar Lidén b
a
IIDEPROQ, UMSA, Plaza del Obelisco 1175, La Paz, Bolivia
b
Department of Chemical Engineering, Lund University, P.O. Box 124, 221 00 Lund, Sweden

Accepted 8 November 2007

Abstract

Quinoa stalk (Chenopodium quinoa Willd.) from agricultural crop residue, totora (Schoenoplectus tatora) and o-macrophytes (aquatic
flora) from Lake Titicaca (on the Bolivian Altiplano) were studied in a wet anaerobic co-digestion process together with manure from
llama, cow and sheep. Anaerobic semi-continuous experiments were performed in (10) 2-l reactors at a temperature of 25 °C with 30 days
of hydraulic retention time (HRT) and an organic loading rate (OLR) of 1.8 kg VS m3 d1.
Totora was found to be the best co-substrate. In mixture ratios of 1:1 (VS basis), it increased the biogas productivity by 130% for
llama manure, 60% for cow manure, and 40% for sheep manure. It was possible to use up to 58% (VS basis) of totora in the substrate.
Higher concentrations (including pure totora) could not be digested, as that caused acidification problems similar to those caused by
other lignocellulosic materials. When quinoa and o-macrophytes were used as co-substrates, the increase in biogas productivity was
slightly less. However, these co-substrates did not cause any operational problems. An additional advantage of quinoa and o-macro-
phytes was that they could be used in any proportion (even in pure form) without causing any destabilization problems in the anaerobic
digestion process.
Ó 2007 Elsevier Ltd. All rights reserved.

1. Introduction The benefits of co-digestion for the anaerobic digestion

Co-digestion, i.e., the simultaneous digestion of a com-
bination of two or more substrates, is an attractive tech-
nique by which the complementary characteristics of the
substrates improve the digestion process. The anaerobic
co-digestion of various agricultural residues (Tafdrup,
1995), various municipal solid wastes (Ahring et al., 1992;
De Baere, 2000), and various other specific organic wastes
(Kaparaju and Rintala, 2005; Mshandete et al., 2004;
Rosenwinkel and Meyer, 1999) is becoming increasingly
attractive as a technique for producing energy in the form
of biogas. It also produces a sludge that is rich in nutrients
and which is often usable as a fertilizer after composting or
other treatment.
* mophilic conditions have been subject to study and com-
Corresponding author. Tel.: +591 46 46 222 8278; fax: +591 46 46
149156.
E-mail address: Rene.alvarez@iideproq.org (R. Alvarez).
process lies in such effects as improved nutrient balance,
improved buffer capacity and improved rheological quali-
ties of the substrate (Sosnowski et al., 2003; Molnar and
Bartha, 1988; Mshandete et al., 2004). There are also
potential economic advantages, such as the possibilities
of utilizing free capacity in digesters in sewage treatment
plants, and of using energy crops as co-substrates (Ahring
et al., 1992; Tafdrup, 1995).
Mixtures of substrates in co-digestion processes have
been shown to be more productive in anaerobic digestion
processes than pure substrates (Mata-Alvarez et al.,
2000). However, negative results have been obtained,
which have been attributed to the specific characteristics
of the co-digested wastes (Murto et al., 2004; Banks and
Humphrey, 1998). Co-digestion under mesophilic and ther-
parisons (Salminen and Rintala, 1999; Kaparaju and
Rintala, 2005). However, there is scarce information about

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Please cite this article in press as: Alvarez, R., Lidén, G., Anaerobic co-digestion of aquatic flora and quinoa with manures ..., Waste
Management (2007), doi:10.1016/j.wasman.2007.11.002
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2 R. Alvarez, G. Lidén / Waste Management xxx (2007) xxx–xxx
co-digestion at low temperatures, about co-digestion of
aquatic flora (macrophytes) or agricultural crop residues
(e.g., quinoa) with manures, for which, there are no previ-
ously published studies.
The Bolivian Altiplano, is a high plateau made up of a
series of plains, highlands, and lakes (Lake Titicaca, Uru-
Uru and Poopo) and situated in the Andes from about
14° to 22° south latitude, at an elevation of more than
3600 m above sea level. The climate is characterized by sea-
sonally low temperatures and intense solar radiation. At
this elevation, less than 5% of the land is suitable for culti-
vation (San Martin and Bryant, 1989); the remaining por-
tion is pastoral and used as pasture for livestock
production. The family herds on the arid Bolivian high-
lands are usually composed of llamas and sheep that feed
exclusively on native range plants (Tichit and Genin,
1997). Quinoa (Chenopodium quinoa Willd.), the ‘‘mother
grain” of the Incas, is remarkably well adapted to the high
altitudes, the arid and saline soils, and to the frequent
frosts that characterize the Altiplano (Prado et al., 2000).
Quinoa grain has an excellent balance of carbohydrates,
lipids, and proteins and provides an ideal mixture of essen-
tial amino acids for human nutrition (Chauhan et al.,
1992). The protein content of quinoa varies between 7.5%
and 22.1% (Tapia et al., 1979). The most important aquatic
vegetation from Lake Titicaca and Uru-Uru Lake are:
totora (Schoenoplectus tatora), llachu (Elodea potamog-
eton), Chancu (Myriophylum ssp.), purima (chara ssp.)
and water lentil (Lemna gibba). The macrophytes occupy
a shallow sector of the two lakes; 15,909 ha distributed
along the Desaguadero River, Lake Titicaca and Uru-
Uru Lake (ABTEMA-UOB, 2000). Llachu and totora con-
stitute the best forage for dairy cattle in the zone; their
digestibility is above 65% and the protein content is above
10% (Quiroz et al., 1997).
In the present work, wet (De Baere, 2000) and low-tem-
perature anaerobic co-digestion was studied as a potential
means to improve the biogas production from manure
from the Bolivian Altiplano. In particular, the effects of
mixing available feedstocks were evaluated in semi-contin-
uous laboratory-scale experiments at 25 °C. The feedstocks
considered were quinoa stalks (residues from quinoa har-
vest), totora and o-macrophytes from Lake Titicaca in
combination with manure (llama, cow and sheep). The
experiments were analysed with respect to gas production,
methane yield and reduction of volatile solids in the
substrates.
Table 1
Characteristics of the fresh undiluted feedstocks used in experiments
Analysis Llama manure Cow manure
Total solids (% w.w) 67.0 16.6
Volatile solids (% of TS) 66.1 75.8
Total Nitrogen (% of TS) 1.9 1.8
Total Organic carbon (% of TS) 28.3 36.1
Total Phosphorous (% of TS) 0.7 0.6
Total Potasium (% of TS) 1.1 1.1
Please cite this article in press as: Alvarez, R., Lidén, G., Anaerobic co-digestion of aquatic flora and quinoa with manures ..., Waste
Management (2007), doi:10.1016/j.wasman.2007.11.002
2. Materials and methods
2.1. Raw materials
Llama, sheep and cow manure were collected from farms
on the Bolivian Altiplano. The manures were separately
ground with a semi-industrial cutter (CUT-3, Metvisa, Bra-
zil), packed in polyethylene bags and stored in a freezer.
The macrophytes, the principal aquatic vegetation in
Lake Titicaca and Uru-Uru–Poopo Lake, were collected
from Lake Titicaca and separated into two groups: totora
(S. tatora) and other macrophytes. The other macrophytes
(o-macrophytes) were principally llachu (E. potamogeton)
and a minor proportion of chancu (Myriophyllum ssp.),
purima (Chara ssp.), and other species such as water lentil
(L. gibba). The totora plants and the o-macrophytes were
separately chopped into smaller pieces (1–4 cm), ground
and homogenized as slurry. The samples were packed into
polyethylene bags and stored at 10 °C in a freezer.
Quinoa (C. quinoa Willd.) crop residue was collected
from farms on the Altiplano (17°S Latitude, 68°W longi-
tude, at an altitude of 3800 m above mean sea level) after
threshing. The samples were cut into 1–4 cm pieces for fur-
ther chopping in a cutting mill. The samples were packed
into polyethylene bags and stored in a freezer.
The materials were analysed for total solids (TS), vola-
tile solids (VS), Nitrogen Kjeldahl, total organic carbon,
potassium, and phosphorus. The composition of the feed-
stocks is given in Table 1.
The inoculum used to seed the reactors to be used for
the experiment was obtained from an active bench-scale
biogas digester working with a mixture of llama–cow–
sheep manure at 30 days HRT and at 25 °C (ST of 9.66%
and VS of 56.51%, pH 7.4).
2.2. Experimental set-up
The anaerobic digestion experiments were carried out in
10 identical stainless steel digesters, each with a total vol-
ume of 2 l. For gas collection, the cylindrical vessels were
equipped with a flanged top to which a flange plate with
stopper ports was fitted. Each digester was equipped with
a port for feeding and effluence drawn through a
12.7 mm ball valve placed close to the base of the reactor.
The temperature was kept constant at 25 ± 1 °C by immer-
sion in a water bath. Two immersion thermostats in each
bath controlled the temperature of the water. Gas collec-
Sheep manure Quinoa Totora o-Macrophytes
59.6 91.9 27.1 7.1
76.9 95.3 89.6 79.7
1.4 2.0 0.8 1.9
22.9 47.8 50.8 42.8
0.5 0.1 0.1 0.2
1.8 1.6 2.3 2.2
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R. Alvarez, G. Lidén / Waste Management xxx (2007) xxx–xxx 3

tion from the reactor was achieved via a flexible PVC tube
to a separate water displacement glass bottle filled with
water acidified to pH 2. The positive pressure in the bottles
allowed the gas to be transferred to a measuring gas cylin-
der. The gas production was recorded at 24-h intervals.
2.3. Experimental procedure
Llama, sheep and cow manure, totora, other macro-
phytes (o-macrophytes), and quinoa were taken out of
the freezer and allowed to thaw overnight. Batches of feed-
stock were prepared according to Table 2. The substrates
were weighed and diluted with tap water, homogenized
with a domestic electric blender, fractionated (volume
defined by the hydraulic residence time, HRT), and packed
into polyethylene bags and stored in a freezer. Every even-
ing, the samples to be used the next day were withdrawn
from the freezer and allowed to thaw overnight.
Anaerobic wet co-digestion experiments with organic
residues from the Bolivian Altiplano were performed in a
semi-continuous process at 25 °C. Llama, cow and sheep
manure were co-digested with quinoa, totora, and o-mac-
rophytes in different proportions. Ten digesters were used
in parallel. The first 10 experiments were started as batch
processes that ran for 20 days before daily feeding was
commenced.
Each reactor was charged with 1800 g of substrate con-
sisting of: inoculum (10%), llama manure (4%), cow man-
ure (4%), sheep manure (14%), and water (68%). The
inoculum from also wet anaerobic digestion process had
9.66% ST, 56.51% SV, and a pH of 7.4. The reactors were
placed in the thermostat controlled water bath at 25 °C,
and allowed to ferment as a batch process for 20 days.
On day 20, the daily feeding was started according to Table
2 with a HRT of 30 days and an organic loading rate
(OLR) of 1.8 kg VS m3 d1. The reactors were maintained
for a period of two HRTs, after that, 10 new feedstocks
were started, using the last active substrate as initial sub-
strate. This procedure was followed until all of the batches
of prepared feedstock were finished. In six experiments
(exp. 1–6) the digesters were fed with unmixed feedstocks
(llama, cow, sheep manure, quinoa, totora, and a mixture
of o-macrophytes), in 12 experiments (exp. 7–18) mixtures
with two components were used as feedstock, and in 7
experiments (exp. 19–25) mixtures with six components
were used in different proportions as shown in Table 2.
The atmospheric pressure during the experiments was
495 mm Hg (the mean atmospheric pressure in La Paz,
Bolivia). The contents of the reactors were mixed by shak-
ing the reactor by hand once a day for about 2 min before
feeding. Once a day, 60 ml of the slurry was withdrawn
from each reactor and replaced with the same volume of
fresh feedstock via the sample removal port. The pH value
and the solid content of the slurry samples were analysed.
Biogas was collected and measured by displacement of
water once a day at zero gauge pressure and at ambient
temperature. The volumes were readjusted to normal
temperature and pressure conditions (0 °C, 760 mm Hg).

Table 2
Experimental design
Exp. no. Llama manure Cow manure Sheep manure Quinoa Totora o-Macrophytes Water
(%VS) (% mass) (%VS) (% mass) (%VS) (% mass) (%VS) (% mass) (%VS) (% mass) (%VS) (% mass) (%VS) (% mass)
1 100.0 11.6 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 88.4
2 0.0 0.0 100.0 47.6 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 52.4
3 0.0 0.0 0.0 0.0 100.0 13.1 0.0 0.0 0.0 0.0 0.0 0.0 0.0 86.9
4 0.0 0.0 0.0 0.0 0.0 0.0 100.0 6.8 0.0 0.0 0.0 0.0 0.0 93.2
5 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 100.0 24.7 0.0 0.0 0.0 75.3
6 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 100.0 98.5 0.0 1.5
7 50.0 5.8 0.0 0.0 0.0 0.0 50.0 3.4 0.0 0.0 0.0 0.0 0.0 90.8
8 50.0 5.8 0.0 0.0 0.0 0.0 0.0 0.0 50.0 12.3 0.0 0.0 0.0 81.8
9 50.0 5.8 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 50.0 49.3 0.0 44.9
10 0.0 0.0 50.0 23.8 0.0 0.0 50.0 3.4 0.0 0.0 0.0 0.0 0.0 72.8
11 0.0 0.0 50.0 23.8 0.0 0.0 0.0 0.0 50.0 12.3 0.0 0.0 0.0 63.8
12 0.0 0.0 50.0 23.8 0.0 0.0 0.0 0.0 0.0 0.0 50.0 49.3 0.0 25.9
13 0.0 0.0 0.0 0.0 50.0 6.5 50.0 3.4 0.0 0.0 0.0 0.0 0.0 90.0
14 0.0 0.0 0.0 0.0 50.0 6.5 0.0 0.0 50.0 12.3 0.0 0.0 0.0 81.1
15 0.0 0.0 0.0 0.0 50.0 6.5 0.0 0.0 0.0 0.0 50.0 49.3 0.0 44.2
16 0.0 0.0 0.0 0.0 0.0 0.0 50.0 3.4 50.0 12.3 0.0 0.0 0.0 84.2
17 0.0 0.0 0.0 0.0 0.0 0.0 50.0 3.4 0.0 0.0 50.0 49.3 0.0 47.3
18 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 50.0 12.3 50.0 49.3 0.0 38.4
19 58.0 6.8 8.0 3.8 8.0 1.0 8.0 0.5 8.0 2.0 8.0 7.9 0.0 78.0
20 8.0 0.9 58.0 27.6 8.0 1.0 8.0 0.5 8.0 2.0 8.0 7.9 0.0 60.0
21 8.0 0.9 8.0 3.8 58.0 7.6 8.0 0.5 8.0 2.0 8.0 7.9 0.0 77.3
22 8.0 0.9 8.0 3.8 8.0 1.0 58.0 4.0 8.0 2.0 8.0 7.9 0.0 80.4
23 8.0 0.9 8.0 3.8 8.0 1.0 8.0 0.5 58.0 14.3 8.0 7.9 0.0 71.5
24 8.0 0.9 8.0 3.8 8.0 1.0 8.0 0.5 8.0 2.0 58.0 57.1 0.0 34.5
25 17.0 2.0 17.0 8.1 17.0 2.2 17.0 1.2 17.0 4.2 17.0 16.7 0.0 65.6
Proportion of substrate (volatile solids and mass percentage basis) in co-digestion experiments.

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Measurements taken over a period of 5 days were averaged
to obtain a final value of the productivity and methane
concentration at steady state. The % VS reduction was cal-
culated according to:
% VS reduction
¼ 100ðVS influent  VS effluentÞ=VS influent
2.4. Analytical methods
The concentrations of methane and carbon dioxide in
the biogas were measured using a gas chromatograph (Shi-
madzu Model GC14B, Japan) equipped with a thermal
conductivity detector (TCD) and a Carboxen-1010 plot
Capillary column 0.32 mm ID (Supelco, USA). Helium
served as the carrier gas. The injector, detector and oven
temperatures were 130 °C, 200 °C and 100 °C, respectively.
The TS, VS, pH, total nitrogen and total phosphorus, in
nitrogen for microbial synthesis; thus the digestibility of
coarse bunch-grasses is improved (Hinderer and Engel-
hardt, 1975; Van Engelhardt and Schneider, 1997).
The characteristics of quinoa stalks, totora and o-mac-
rophytes are given in Table 1.The TS value of 89.4% in qui-
noa stalks is obtained from stalks dried outdoors. This
practice of drying reduces the humidity of the grain from
25% to 10% in order to facilitate threshing (separation of
the grain) (Tapia et al., 1979). The carbon/nitrogen (C/
N) ratio of 22.7 shows an excellent balance of organic mat-
ter to nitrogen for optimal digestion in an anaerobic diges-
tion process (NAS, 2001).
The aquatic vegetation in Lake Titicaca was represented
by totora (S. tatora) and a mixture of other macrophytes.
The mixture of other macrophytes was composed of llachu
(E. potamogeton), chancu (Myriophyllum ssp.), purima
(Chara ssp.), and water lentil (L. gibba). Table 1 shows

Biogas Productivity (ml d-1)

the feedstocks and in the substrate samples, were all ana- 1500
a
lysed using standard methods as described by APHA
(2000). The TS content was determined after a procedure 1000
of heating (105 °C for 1 h), cooling, desiccating, and weigh-
ing repeated until constant weight was reached. VS were 500
determined in a muffle furnace at a temperature of
550 °C by ignition of the residue produced during the TS 0
analysis to constant weight. Total Kjeldahl nitrogen
(TKN) was measured using the semi-micro-Kjeldahl
Biogas Productivity (ml d-1)

1500
method as described in Standard Methods (APHA, b
2000). Potassium and phosphorus were measured by spec-
trophotometry (Method 3500-K and 4500-P, respectively). 1000

500
3. Results and discussion

3.1. Raw material 0

Biogas Productivity (ml d-1)

The composition of the raw material used in the exper- 1500

iments are shown in Table 1. The composition of the man- c
ures obtained from the Altiplano agreed well with the 1000
reported mean values for beef and sheep manure (ASAE,
2003) The exception was the nitrogen content in the cow 500
manure, which was low: 1.6% of TS, i.e., less than 41%
of the values reported by ASAE (2003). This probably
0
reflects the low digestibility and nutritional value of the
pasturage and the difficulty of providing cultivated forage
Biogas Productivity (ml d-1)

to supplement the diet of farm livestock on the Altiplano 1500

(Quiroga, 1992; Alzerreca, 1992). The nitrogen and solids d
contents of the llama manure are the highest of all consid- 1000
ered manures, and among the highest reported in ASAE
(2003). A study conducted on the nitrogen requirements 500
of South American camelids, estimated that their mainte-
nance requirements are lower than those of sheep and beef 0
cattle (Preston, 1996). In addition, the higher digestibility
0 20 406 0
value in llamas on low nitrogen forage is explained by
the llama’s ability to maintain higher NHþ 4 concentrations
Time (d)
in the first and second compartments of the stomach com- Fig. 1. Biogas productivity from anaerobic digestion of llama manure (a),
pared to sheep. The llamas are hereby provided with more quinoa (b), totora (c) and other macrophytes (d).

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the major differences between totora and the mixture of
other macrophytes in total solids (27% and 7%), nitrogen
(0.8% and 1.9%), and organic carbon (56% and 35%,
respectively). The low solids content of the mixture of other
macrophytes may impose a serious practical restriction on
its use in wet anaerobic digestion as carbon source; how-
ever. It can be used as a source of water.
The C/N ratio is 63.5 in totora and 22.5 in the mixture
of other macrophytes. In principle, the high C/N ratio in
totora would make it inadequate for an optimal digestion
process. Nevertheless, in practice overall C/N values can
vary considerably (from less than 10 to over 90) without
affecting the efficiency of digestion (NAS, 2001; Marchaim,
1992) as this is due to the fact that not all the carbon and
nitrogen in the feedstock are available for anaerobic diges-
tion. The biodegradable (real) C/N ratios are lower than
the total C/N, which is based on a total C determination
(Sánchez, 2007). Biodegradable organic carbon is a func-
tion of the type of feedstock and operational process; the
amount of organic carbon to be aerobically degraded is sig-
nificantly higher than the anaerobic biodegradable fraction
(Komilis and Ham, 2003).
3.2. Co-digestion experiments
The co-digestion of organic wastes depends on the rela-
tive proportions of the components (on a certain basis such
as weight, volume or VS content), the amount of the mix-
ture and other physical process parameters, such as tem-
perature and pressure. In the current experimental design,
the independent factors are the fractions of the six compo-
nents of a blend. As the fractions of the different compo-
nents always sum to 1 (100% total of volatile solids), the
total VS content was considered to be the parameter to
normalize. The same HRT, OLR, and temperature were
maintained in all experiments. The atmospheric pressure
was 495 mm Hg. Consequently, the responses or dependent
factors (biogas productivity, methane yield and percentage
VS removed) depend only on the substrate composition.
The simplex-centroid design for the mixture of six compo-
nents was used as the base in the current work (Khuri and
Cornell, 1996; Montgomery, 2001), giving the experiments
in Table 2.
In general, the evolution of the anaerobic digestion
experiments using unmixed feedstock (llama, cow and
sheep manures, quinoa and o-macrophytes) showed a sta-
ble pattern after an adaptation period of about 30 days
(1 HRT); the pH-values ranged between 6.9 and 7.2 with
a variation of less than half a unit within a single experi-
ment (Fig. 1). Biogas productivity reached steady states
in the range of 400–850 ml d1 with a methane content
between 49% and 55% and methane yields of 0.07–
0.14 m3 kg1 VS added (Table 3). Experiment 5 (digestion
of totora) was the exception to the general pattern

Table 3
Proportion of substrate and steady-state results from anaerobic co-digestion of agricultural residues, aquatic flora and manure
Exp. Proportion of substrate Effluent VS Biogas Methane Methane yield
no. pH reduction productivity content (%) (m3 kg1 VS added)
Llama Cow Sheep Quinua Totora o-
Macrophytes (%) (ml d1)

1 1.00 0.00 0.00 0.00 0.00 0.00 7.2 19 416 (34) 53 (2) 0.067 (0.005)
2 0.00 1.00 0.00 0.00 0.00 0.00 7.1 14 579 (46) 54 (3) 0.094 (0.007)
3 0.00 0.00 1.00 0.00 0.00 0.00 7.1 17 574 (14) 53 (3) 0.092 (0.002)
4 0.00 0.00 0.00 1.00 0.00 0.00 7.1 32 535 (18) 49 (3) 0.079 (0.003)
5 0.00 0.00 0.00 0.00 1.00 0.00 5.0 37 179 (77) 27 (5) 0.015 (0.006)
6 0.00 0.00 0.00 0.00 0.00 1.00 7.0 20 848 (16) 55 (2) 0.141 (0.003)
7 0.50 0.00 0.00 0.50 0.00 0.00 6.9 17 676 (30) 51 (3) 0.104 (0.005)
8 0.50 0.00 0.00 0.00 0.50 0.00 7.1 31 980 (32) 52 (3) 0.154 (0.005)
9 0.50 0.00 0.00 0.00 0.00 0.50 7.3 16 656 (28) 54 (2) 0.107 (0.005)
10 0.00 0.50 0.00 0.50 0.00 0.00 6.9 21 619 (29) 50 (1) 0.093 (0.004)
11 0.00 0.50 0.00 0.00 0.50 0.00 7.2 21 989 (38) 50 (1) 0.149 (0.006)
12 0.00 0.50 0.00 0.00 0.00 0.50 7.1 24 748 (18) 54 (1) 0.122 (0.003)
13 0.00 0.00 0.50 0.50 0.00 0.00 7.1 25 635 (20) 51 (1) 0.098 (0.003)
14 0.00 0.00 0.50 0.00 0.50 0.00 7.1 26 871 (32) 49 (1) 0.129 (0.005)
15 0.00 0.00 0.50 0.00 0.00 0.50 7.3 19 683 (9) 52 (1) 0.108 (0.001)
16 0.00 0.00 0.00 0.50 0.50 0.00 7.2 43 900 (27) 46 (4) 0.124 (0.004)
17 0.00 0.00 0.00 0.50 0.00 0.50 7.3 17 805 (9) 53 (1) 0.128 (0.001)
18 0.00 0.00 0.00 0.00 0.50 0.50 7.0 24 1255 (20) 51 (1) 0.192 (0.003)
19 0.58 0.08 0.08 0.08 0.08 0.08 7.2 21 624 (31) 52 (2) 0.098 (0.005)
20 0.08 0.58 0.08 0.08 0.08 0.08 7.0 15 745 (31) 51 (1) 0.115 (0.005)
21 0.08 0.08 0.58 0.08 0.08 0.08 7.1 21 593 (12) 53 (1) 0.096 (0.002)
22 0.08 0.08 0.08 0.58 0.08 0.08 7.2 27 812 (50) 49 (3) 0.121 (0.007)
23 0.08 0.08 0.08 0.08 0.58 0.08 7.2 37 951 (280) 51 (3) 0.147 (0.043)
24 0.08 0.08 0.08 0.08 0.08 0.58 7.4 33 927 (23) 50 (1) 0.140 (0.004)
25 0.17 0.17 0.17 0.17 0.17 0.17 7.1 16 844 (15) 51 (1) 0.130 (0.002)
Standard deviation from 5 consecutive days in parentheses.
o-Macrophytes: llachu (Elodea potamogeton), chancu (Myriophyllum ssp.), purima (Chara ssp.) and water lentil (Lemna gibba).

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6 R. Alvarez, G. Lidén / Waste Management xxx (2007) xxx–xxx

described above; the digestion process showed signs of

Biogas productivity (ml d-1)

1600 a
acidification and the pH dropped slowly from 7.1 (until
day 24) to 5 (day 60); biogas volumes and methane content 1200
also showed declining patterns (Fig. 1c), e.g., the methane
800
content dropped to 25% (day 60) showing signs of
inhibition. 400
The behaviour of totora in exp. 5 showed the typical
problem associated with digestion of lignocelluloses, i.e., 0
the difficulty of maintaining a stable pH within the correct

Biogas productivity (ml d-1)

range (Brummeler and Koster, 1990). This is associated 1600 b
with a poor buffering capacity and the accumulation of vol-
1200
atile fatty acids (Banks and Humphrey, 1998). The co-
digestion of totora and other wastes turned out to be a 800
good attempt to solve the problems associated with pH
(see below). 400
The co-digestion experiments that used mixtures of two 0
co-substrates (exp. 7–18) with 50:50% in volatile solids
basis, reached stable profiles with little fluctuation in pH

Biogas productivity (ml d-1)

and methane content after the adaptation period, which 1600 c
indicates improved methane productivities (co-digestion 1200
effect) (Fig. 2). The steady-state methane yields obtained
were in the range of 0.09–0.2 m3 kg1 VS added, the biogas 800
production was 619–1255 ml d1 with 46–54% of methane
400
content (Table 3).
The methane yields of llama manure increased as a 0
result of co-digestion: by 130% with totora, 110% with qui-
noa and 60% with o-macrophytes. The pH value and the
Biogas productivity (ml d-1)

1600 d
methane content varied little over the course of these exper-
iments. The results show that llama manure benefits by 1200
anaerobic co-digestion with quinoa (agricultural waste),
800
totora and o-macrophytes (aquatic flora).
The co-digestion of cow manure + quinoa and of 400
sheep + quinoa in the ratio 1:1 showed a minor increase
0
in methane yield; compared to a digester working with pure
cow manure, these mixtures produced up to 7% more
Biogas productivity (ml d-1)

methane. However, the co-digestion of cow manure and 1600 e

aquatic flora showed more positive results; the increases
in methane yield were: 59% cow + totora, 30% cow + o- 1200
macrophytes, 40% sheep + totora, and 17% sheep + o- 800
macrophytes.
From the co-digestion experiments using the three man- 400
ures together with quinoa, with totora and with o-macro-
0
phytes in the ratio 1:1 (VS basis), it was concluded that
totora was the best co-substrate, increasing the biogas pro-
Biogas productivity (ml d-1)

ductivity by 130%, 60%, and 40% for llama, cow and sheep, 1600 f
respectively. In co-digestion with quinoa and o-macro- 1200
phytes, the digestion of llama, cow and sheep manure
was slightly less improved and the co-substrates did not 800
cause any operational problem.
400
In our experiments, the anaerobic digestion of mixtures
improved performances compared to digestion of single 0
substrates. However, the highest productivity is achieved 0 20 40 60
by careful incorporation of an adequate proportion of lig- Time (d)
nocellulosic materials. For example, in the co-digestion
Fig. 2. Co-digestion experiments with mixtures of two substrates: llama
with all six components, the digestion of mixtures with a manure with o-macrophytes (a), sheep manure + totora (b), and cow
higher content of totora (58% VS basis, exp. 23) showed manure + quinoa (c), totora + o-macrophytes (d), o-macrophytes + qui-
high biogas production and stable values of pH and meth- noa (e) and quinoa + totora (f).

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R. Alvarez, G. Lidén / Waste Management xxx (2007) xxx–xxx 7

1600 4. Conclusion
Biogas productivity (ml d-1)

a
1200 Agricultural wastes and aquatic flora are good co-sub-
strates for anaerobic co-digestion processes.
800 The results obtained from the co-digestion experiments
showed improved performance as a result of mixing man-
400
ure with crop residues and aquatic biomass. Totora was
0
the best co-substrate.

1600
Biogas productivity (ml d-1)

b Acknowledgement
1200
This work was financially supported by SIDA, the
800 Swedish International Development Cooperation Agency.

400
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