ARTICLE IN PRESS

Biomass and Bioenergy 30 (2006) 66–75

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Biogas production from llama and cow manure at high altitude

René Alvareza,b, Saul Villcaa, Gunnar Lidénb,
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
Received 14 June 2004; received in revised form 9 October 2005; accepted 10 October 2005
Available online 23 November 2005

Abstract

Methane production from llama and cow manures from the Bolivian high plateau (The ‘‘Altiplano’’) was studied using a parallel reactor
set-up consisting of 10 lab-scale biogasifiers. The effects of pressure (495 and 760 mmHg), temperature (11 and 35
C), hydraulic retention
time (20 and 50 days), and manure content in the slurry (10%, 20% and 50%) were evaluated with respect to productivity and methane
yields based on two 241 fractional factorial designs with 8 treatments for each kind of manure. The reactors were operated semi-
continuously with daily manure feeding for periods between 50 and 100 days. Temperature was the main factor effect found, and the
hydraulic retention time and the manure content in feed were also found significant whereas the effect of pressure was not significant in the
range studied. The methane yield obtained with cow manure at 11
C was between 6.4 and 33:6 l CH4 kg1 VS (volatile solids added)
whereas at 35
C the methane yield was between 49.6 and 131:3 l CH4 kg1 VS. The methane yield from llama manure was somewhat lower
than for cow manure (between 3.3 and 19:3 l CH4 kg1 VS at 11
C and between 35.6 and 84:1 l CH4 kg1 VS at 35
C, respectively).
However, overall llama manure was found to be the best raw material of the two for biogas production, due to its high content of volatile
solid—higher than has been previously reported for most manures—and also its high nitrogen and phosphorous content.
r 2005 Elsevier Ltd. All rights reserved.

Keywords: Anaerobic digestion; Cow manure; Llama manure; High altitude

1. Introduction The Andean high plateau (the Altiplano) is a region

Anaerobic digestion of animal wastes for production of
biogas is a widely studied subject. Properly functioning
biogas systems can yield a whole range of benefits for their
users, including production of heat, light, and electricity,
transformation of organic waste into high-quality fertilizer,
improvement of hygienic conditions through reduction of
pathogens, reduction of work for firewood collection and
cooking, and environmental advantages through protec-
tion of soil, water, air, and woody vegetation [1,2].
However, most studies concern biogas production in warm
countries and at sea level. There have been only few studies
on biogas production at intermediate altitudes
(1000–3000 m over the mean sea level), and for highlands
above 3000 m over the sea level there are in fact no
published studies.
Corresponding author. Tel.: +46 46 222 0862.
E-mail address: Gunnar.Liden@chemeng.lth.se (G. Lidén).
covering more than 100; 000 km2 situated between the
southwest of Bolivia and the south of the Peru (15–22
S
latitude, 65–72
W longitude). The altitude of the region is
between 3000 and 4000 m above sea level. The mainly rural
population is still heavily dependent on traditional fuels,
such as firewood, animal wastes and agricultural residues.
The livestock farms hold llamas (currently more than 3
million animals), alpacas, cows, and sheep. The energy
demands give rise to an excessive firewood consumption and
deforestation. There has therefore been an increased interest
in the development of technologies for harnessing renewable
energy sources, such as biomass, either directly or through
conversion routes. Of the biological processes, anaerobic
digestion is a serious option [1,2]. Biogas production at the
Altiplano is, however, associated with a number of unique
characteristics with respect to pressure, temperature and
feedstock characteristics, which need to be understood.
The production of methane from anaerobic digestion of
livestock manures is known to depend on the kind of

0961-9534/$ - see front matter r 2005 Elsevier Ltd. All rights reserved.
doi:10.1016/j.biombioe.2005.10.001
 ARTICLE IN PRESS
R. Alvarez et al. / Biomass and Bioenergy 30 (2006) 66–75
material added to the digester, the solids loading, the
temperature, and the hydraulic retention time [2,3]. As to be
expected, a strong temperature effect has been observed by
many researchers [3–5], and the biomethanation process at
mesophilic and thermophilic ranges is relatively well under-
stood and documented. However, concerning the anaerobic
digestion at psychrophilic temperature there is a severe lack
of fundamental knowledge [6]. With respect to retention
time, the fermentation has been report to become more
stable with a higher methane yield and reduction of VS with
an increasing hydraulic residence time (HRT) [3]. Long
retention times are required for manure digestion, not only
due to the presence of complex organic compounds, but also
due to the high concentration of ammonia nitrogen, which
affects anaerobic decomposition [7]. The current practice in
unheated biogas plant in, for example, India is to use a
retention time of about 30 days in warm climates and up to
55 days in the colder hilly regions [8].
The effect of increasing the pressure in the anaerobic
digestion has been addressed by several researchers. The
prime effect relates to the increasing carbon dioxide
concentration in the liquid phase caused by an increased
pressure. Hayes et al. [9] reports an increasing methane
content in a digester with higher pressure. Carbon dioxide
is 40 times more soluble in water than methane, and a high
digester pressure therefore results in high concentration of
carbon dioxide in the substrate, which drives the methane
producing reactions forward and stimulates methane
production. By changing the total gas pressure of the
anaerobic digester toxicity effects can be avoided. An
increase of CO2 partial pressure decreases the pH value,
which reduces the non-ionized ammonia concentration. On
the other hand, a decrease in CO2 partial pressure increases
the pH level, which lowers the non-ionized hydrogen-
sulfide concentration [10]. The pressure created during
anaerobic digestion has been used as the source of mixing
power. The mixing was achieved through gas recirculation,
utilizing the pressure of gas in the storage tank as a power
source. The average gas production rate observed was
comparable to that of a commercial digester with mechan-
ical agitation [11].
Comparison of different animal manures as a digester
feedstock for local distributed biogas generation has been
made for typical low-land farms. Using dairy and swine
manure, Safley and Westerman [4] estimated that the
methane yield increased almost linearly over the tempera-
ture range of 10–23
C. Higher yield was suggested for the
lower loading rates (LRs) (0.1 kg VS m3 d1 ) compared
with higher (LRs) (0.2 kg VS m3 d1 ). Hammad et al. [12]
compared cattle, poultry, sheep and horse manure for
methane production, and obtained the highest productivity
of biogas with cattle manure (0:28 m3 m3 d1 at 35
C).
Axaopoulos et al. [13], improved the productivity of a swine
manure digester by using solar heating, and obtained an
average daily methane production rate of 0:64 m3 CH4 m3
digester. Kalia and Singh [14] studied mixtures of horse and
cattle dung in different ratios for running family-size biogas
67
plant biomethanation in a hilly region and reported a biogas
production of 0.325–0:75 m3 m3 d1 . It was found that a
20% substitution of cattle dung with horse dung could be
made without much reduction in the daily gas production.
The biogasification related to the seasonal variation in
the ambient temperature on highlands was evaluated by
Kalia and Kanwar [15] in a Janata biogas plant installed in
hilly conditions (1300 m above the sea level). The monthly
variation in the mean ambient temperature (9–26
C)
affected its digester temperature (13–23
C) and, hence,
rate of gas production (0.26–0:5 m3 m3 d1 with cow
manure). Kanwar and Guleri [16] compared the perfor-
mance of a rubber-balloon biogas plant with a fixed-dome
plant at 1300 m elevation. In the rubber-balloon plant, the
average daily rate of biogas production during summer
(16–24
C) was 0:03 m3 kg1 feed but only 0:007 m3 kg1
feed in the winter (13–15
C), showing the high effect of the
temperature.
The objective of the current work was to study the
production of biogas from typical manure sources of the
Altiplano, i.e. cattle and llama manure. A parallel reactor
set-up consisting of 10 lab-scale biogasifiers was used, and
the reactors were operated semi-continuously with daily
manure feeding for periods between 50 and 100 days. In
addition to the feedstock, also temperature and pressure as
well as the hydraulic retention time and solids loading were
studied using a fractional factorial designs. The experi-
ments were statistically evaluated and analysed with
respect to productivity and methane yields.
2. Materials and methods
2.1. Feedstock
Llama and dairy cattle manure were collected from
farms in the Bolivian Altiplano situated at 19
S Latitude
and 68
W longitude at an altitude of 3800 m above mean
sea level. The samples were packed into 500 g polyethylene
bags and stored at 10
C in a freezer until used.
2.2. Apparatus
The experiments were carried out in a semi-continuous
process using ten digesters, each with a working volume of
2 l. The digesters were built based on the design used by
Chowdhury and Fulford [17]. Each digester was equipped
with inlet and outlet ports for feeding and effluent
discharge, respectively, and with a port for collecting gas
(Fig. 1). The gas measuring cylinder, the digesters and the
collecting biogas bottle were all made of glass. The water
bath operating at low temperature ð11
CÞ was controlled
by a system with two immersion Pt100 type sensors and an
analog temperature controller (Pico, Pico technology
limited, UK). The water bath operating at 35
C was
controlled by an immersion thermostat.
The pressure in the digester and the gas collecting bottle
was set by the difference in water level between the water
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68 R. Alvarez et al. / Biomass and Bioenergy 30 (2006) 66–75
Water displacement bottle
Gas measuring cylinder
Water reservoir
Water bath
Gas collection bottle Digester
Fig. 1. Schematic diagram of the anaerobic digestion system.
displacement bottle and the gas collecting bottle. The
average atmospheric pressure in La Paz City is 495 mmHg,
and to obtain a digester pressure of 760 mmHg a water
level difference of 360 cm was thus needed.
The biogas was collected in the collecting bottles, acting
as gas reservoirs. The overpressure in the bottles allowed
the gas to be transferred to a measuring gas cylinder and
this measurement was made once a day. The feed of fresh
substrate and the removal of digested material were made
carefully in order to maintain the pressure difference in
each digester.
2.3. Experimental procedure
Llama and dairy cattle manure were withdrawn from the
freezer and allowed to thaw overnight. The slurry was
diluted with tap water to obtain the desired solid-content
and the slurry was then homogenized with a domestic
electric blender (Hamilton Beach 908, Hamilton Beach
Commercial, USA).
Ten semi-continuous experiments were made in parallel
(five fermentors at 11
C and five at 35
C). The experi-
ments were started as batch processes, which ran for 15
days before daily feeding was commenced. A volume of
slurry (defined by the value of the desired HRT) was
withdrawn from each reactor daily and was replaced with
the same volume of fresh feedstock, via the slurry sampling
tube. All reactors were mixed by shaking the glass flasks by
hand once a day for about 2 min. This mimicked the
practice adopted for rural digesters, which are normally
mixed only once a day.
The establishment of a steady state in the reactors was
assessed by measuring the daily gas production and the
methane content of the gas, as well as the pH. When a
reactor had reached a stable condition (usually after about
2 HRT), measurements taken for a period of several days
were averaged to obtain a final value for the productivity
and methane concentrations.
2.4. Analytical methods
The gas volume was measured at zero gauge pressure
and ambient temperature. The biogas was analysed for
methane content and the volume corrected to dry gas at
standard temperature and pressure (0
C and 760 mmHg).
Methane and carbon dioxide concentrations in the biogas
were determined with a gas chromatograph (Shimadzu
Model GC14B, Japan) equipped with a thermal conduc-
tivity detector (TCD) and Carboxen-1010 plot Capillary
column 30 m  0:53 mm ID (Supelco, USA). The injector,
detector and oven temperatures were 150, 200 and 120
C,
respectively. Helium served as the carrier gas at a pressure
of 300 kPa.
Total solids (TS), fixed solids (FS) and VS, pH, oil-
grease, and alkalinity were determined according to
standard methods [18]. The TS content was determined
after a repeated heating (105
C for 1 h), cooling, desiccat-
ing, and weighing procedure until the weight change was
less than 4%. FS and VS were determined by ignition of
the residue produced in TS analysis to constant weight in a
muffle furnace at a temperature of 550
C. The oil and
grease content was analysed by measuring the amount of
material, extractable with n-hexane in a Soxhlet extraction
instrument after hydrolysis with concentrated HCl. The
alkalinity was determined by a titration method [18].
Chemical oxygen demand (COD), total Kjeldahl nitro-
gen (TKN), ammonia, and phosphorus were measured
with Hach analysis systems (Hach Company, Loveland,
Colorado, USA) composed of a spectrophotometer (DR/
4000 UV VIS), COD Reactor (45600-00), COD digestion
reagent vials (0–15,000 mg/l COD), total nitrogen vials
(10–150 mg/l N), total ammonia vials (0–50 mg/l NH3 –N),
and total phosphorus (0–3.5 mg/l PO3 4 ) according to the
procedures from the manufacturer.
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R. Alvarez et al. / Biomass and Bioenergy 30 (2006) 66–75 69

2.5. Factorial design Table 2

Composition of the fresh undiluted manures
Four different factors for the production of biogas Type of analysis Cow Llama
from llama and cow manure of the Bolivian Altiplano
were studied, i.e. pressure, temperature, retention time, and Total solids (% wet) 15.6 59.5
solids loading (percentage of dung in the slurry). The Volatile solids (% TS) 82.8 74.4
Fixed solids (% TS) 17.2 25.6
experiments were set-up as two 241 fractional factorial
Total nitrogen (% TS) 1.2 1.9
designs [19]. There were thus 8 treatments for each kind Nitrogen ammonium (% 0.05 0.14
of manure. TS)
The factor levels, given in Table 1, were chosen based on Phosphorus (g/kg) 2.4 5.2
the feasible operative range for the anaerobic digestion of Fe total (g/kg) 3.9 2.4
Oil and greasy (% TS) 81.2 36.7
the agricultural wastes. pH 7.8 7.8

3. Results 3.2. Biogas production

3.1. Characteristics of feedstock The biogas productivities, methane content in the gas at
steady state as well as the feeding conditions are
The cow and llama manures were analysed with respect to summarized in Table 3. The steady-state values of
solids and VS content, nitrogen and ammonia nitrogen productivity and yield were based on an average of
content, phosphorous content, and oil and grease content measurements over a 6-day period after a minimum of 2
(Table 2). There were several important differences between retention times. The daily biogas production (measured at
the two sources of manure. Particularly noticeable was the local conditions, i.e. 495 mmHg and 15
C) was rather
much higher solid content in llama manure, more than 3 different at the different experimental conditions. The
times that in cow manure. The unusual high content of biogas production was between 90 and 1010 ml d1 when
solids in llama manure was compared with previously using cow manure as feedstock and between 70 and
published data for cow, pig, or horse faeces [2,14,20]. The 1100 ml d1 with llama manure.
solids content of llama manure was more than two times The methane content in the biogas varied from about
higher than the reported values of those manures. The total 40% to 60% with cow manure and 20% to 57% with llama
nitrogen content in the cow manure was low, less than 50% manure. The measured daily biogas production and the
of the reported value from dairy cattle [2,20]. This most methane content in the gas in four representative experi-
likely reflects the feed situation on the Altiplano. Further- ments are shown in Fig. 2. The chosen experiments
more, the low ammonia content in the cow manure can be correspond to exp. no. 12 and 14 (cow manure) and exp.
noted (less than 10% of reported values in [2,20]). This may no. 9 and 15 (llama manure) (cf. Table 3).
be attributed to the feed, as well as a fast loss of ammonia by Table 4 show the factorial design with 8 experiments for
volatilisation at low humidity and windy conditions on the each kind of manure, and the measured primary response
Bolivian Altiplano. in terms of volumetric productivity ð1 day1 Þ and methane
yield ð1 CH4 kg1 VSÞ. A logarithmic transformation was
made to stabilize the variance. The analysis and signifi-
cance of the transformed variables are summarized in
Table 1 Table 5.
Factors and levels for the 241
IV fractional factorial design used for cow and The analysis of variance showed that the main effects
llama manure digestion
dominate, explaining more than 92% of the total
Factors Levels variability, whereas the interaction effects explain less than
8% of the total variability. The temperature effect was
1 1
significant in all cases. As expected, the effect of the
Cow manure temperature was positive, and an increment in the
A: Pressure (mmHg) 495 760 temperature produces a considerable increase in the daily
B: Temperature ð
CÞ 11 35 production of biogas and it increases the yield of methane.
C: Retention time HRT (days) 20 50
Perhaps more suprisingly, however the pressure effect was
D: Dung in slurry (%) 20 50
not found to be significant in any case. The other main
Llama manure factor effects, i.e. residence time and loading of solids in the
A: Pressure (mmHg) 495 760 feed, were found to be significant for the methane yield and
B: Temperature ð
CÞ 11 35 the biogas productivity, respectively.
C: Retention time HRT (days) 20 50
The HRT (C) was the second effect in magnitude to the
D: Dung in slurry (%) 10 20
sum of squares (16.36% in llama and 20.1% in cow) when
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70 R. Alvarez et al. / Biomass and Bioenergy 30 (2006) 66–75

Table 3
Biogas productivities and methane contents obtained at different loading rates

Trial Manure Total solid Volatile solid Loading rate Biogasa Methane content
no. ðkg d1 Þ ðkg TS d1 Þ ðkg VS d1 Þ ðkg VS m3 d1 Þ production rate (%)
ðml d1 Þ

Cow manure
2 0.0180 0.0028 0.0023 1.29 102  19 46  8
4 0.0450 0.0070 0.0058 3.22 141  19 43  1
6 0.0450 0.0070 0.0058 3.22 1010  68 46  8
8 0.0180 0.0028 0.0023 1.29 479  40 54  8
10 0.0180 0.0028 0.0023 1.29 226  34 56  6
12 0.0072 0.0011 0.0009 0.52 93  31 39  6
14 0.0072 0.0011 0.0009 0.52 324  15 61  4
16 0.0180 0.0028 0.0023 1.29 475  43 57  4

Llama manure
1 0.0180 0.0107 0.0080 4.43 202  29 21  3
3 0.0090 0.0054 0.0040 2.22 68  23 47  7
5 0.0090 0.0054 0.0040 2.22 401  61 57  13
7 0.0180 0.0107 0.0080 4.43 1096  84 52  4
9 0.0036 0.0021 0.0016 0.89 87  8 57  4
11 0.0072 0.0043 0.0032 1.77 128  43 46  6
13 0.0072 0.0043 0.0032 1.77 457  35 49  8
15 0.0036 0.0021 0.0016 0.89 519  39 42  4
a
Daily volume in local conditions (P ¼ 495 mmHg, T ¼ 15
C).

800 100 13.74% using the llama and cow manures, respectively.
The effect of the manure content in slurry was positive.
Biogas volume (ml d )

80
-1

Methane content (%)

600 The results can be visualized in an different way by

60 plotting the production rate of methane ðml d1 Þ and
400 the methane yield ð1 kg1 VSÞ vs. the LR ðgVS L1 d1 Þ
40
(Figs. 3 and 4). Apparently, the productivity increases
200
20 continuously, whereas the methane yield decreases con-
tinuously in the range studied.
(a) 0 0

4. Discussion
800 80

4.1. Comparison of feedstocks

Methane content (%)
Biogas volume (ml d )
-1

600 60

Due to the higher content of TS in llama manure

400 40
(Table 2), llama manure has a higher content of VS,
200 20
nitrogen, and phosphorous, per unit of wet manure weight
than cow manure. Also in comparison to other previously
0 0 reported values of (Table 6) wet llama manure has a very
1 10 25 40 55 70 85 100 115
high content of VS, nitrogen, and phosphorous in
(b) Days
comparison to dairy cattle, beef cattle, swine, sheep, goat,
Fig. 2. Volumetric biogas productivity in semi-continuous anaerobic poultry, and horses. The nitrogen content in the cow
digestion of (a) cow manure and (b) llama manure at 11
C ð’Þ and manure on the other hand was surprisingly low compared
35
C ðEÞ. Also the methane content is shown at 11
C ð&Þ and to other reported values for cattle (Table 6). The content of
35
C ðBÞ.
nitrogen in the cow manure from the Bolivian highland is
in fact less than the reported minimum requirement (0.6%)
needed for incorporation into the methanogenic bacteria
considering the methane yield. The effect of the HRT was cell structure, and it is therefore possible that the gas
positive implying increase in the methane production when production will be limited by the nitrogen availability [2].
increasing the time of retention, which agrees with other This limitation could be one of the reasons for the observed
reports [3,8]. The manure content in slurry (D) was low production of biogas from cow manure obtained in the
significant when considering the biogas productivity, with present work. On the other hand, an excess of nitrogen in
a contribution to the sum of squares of 10.49% and the substrate leads to ammonia formation, which may
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R. Alvarez et al. / Biomass and Bioenergy 30 (2006) 66–75 71

Table 4
Experimental design and measured responses

Trial Temperature Pressure HRT Manure Biogas ðnÞa Methane yield

no. ð
CÞ (mmHg) (day) in slurry production ð1 CH4 kg1 VS loadÞ
(%) ðml d1 Þ

Cow manure
2 11 495 20 20 63  12 12:5  2:4
4 11 760 20 50 87  12 6:4  0:9
6 35 495 20 50 623  42 49:6  3:4
8 35 760 20 20 296  25 69:5  5:9
10 11 495 50 50 140  21 33:6  3:6
12 11 760 50 20 58  19 24:4  8:4
14 35 495 50 20 200  9 131:3  6:2
16 35 760 50 50 294  27 72:4  5:4

Llama manure
1 11 495 20 20 125  18 3:3  0:5
3 11 760 20 10 42  14 5:0  1:7
5 35 495 20 10 248  38 35:6  5:4
7 35 760 20 20 677  52 43:9  3:3
9 11 495 50 10 54  5 19:3  1:7
11 11 760 50 20 79  26 11:5  3:8
13 35 495 50 20 282  22 43:4  3:3
15 35 760 50 10 320  24 84:1  6:3
a
Biogas production in normal conditions (760 mmHg, 0
C).

cause inhibition of cell growth and lead to a slower, or even
complete stop of the production of methane [21]. In the
present case, the nitrogen content of the llama manure is
1.13%, i.e. higher than the swine manure (0.52%, Table 6),
but the initial ammonia nitrogen is only 0.083% (7% of the
total nitrogen, Table 6) which decreases the risk for initial
inhibition. The risk of ammonia inhibition will increase
with LR in the digester. For unadapted methanogenic
cultures, ammonia inhibition has been observed to
commence at concentration of between 1.5 and 2:5 g N l1
[22]. However, by adaptation of the biogas process to
ammonia, tolerance to 4 g N l1 total ammonia has been
demonstrated [22,23]. This suggests that the methane yield
obtained in the present work could be improved by
adaptation of the biogas process, limiting the % manure
in the substrate or by mixing of llama manure with other
co-substrates.
4.2. Main factor effects
Based on the design, it was found that the temperature,
the hydraulic retention time and the solids loading (% wet
dung) were significant factors with respect to biogas
productivity and methane yield in the anaerobic digestion
process. Not surprisingly, temperature was found to be the
main factor (Tables 3–5) and the increase of the
temperature from 11 to 35
C dramatically increased both
the biogas productivity (4 and 5 times for cow and llama
manure, respectively) and the methane yield (4 and 5 times
for cow and llama manure, respectively). The effect of
temperature on the production of biogas has been reported
by several investigators [3–5,8,24]. From a very simplistic
kinetic viewpoint, one could argue that the Arrhenius law
would predict an exponential increase in rate as a function
of temperature. However, the situation is far more complex
since many species of microorganisms operate in the
reactor, and different temperatures will lead to a selection
of certain species. Safley and Westerman [4] working with
dairy manure reported an almost linear increase in gas
production with increasing temperatures in the range of
10–23
C. In the present investigation, the temperature
difference is probably too large for such a linear increase to
be valid.
The hydraulic retention time is the second most
important factor found in the present work for the methane
yield. An increased retention time had a positive effect for
both substrates. Similar findings have been reported for
cattle manure [3,8] and solid poultry slaughterhouses
wastes [25].
The percentage of manure in the slurry was also found to
be a significant parameter although somewhat less im-
portant than temperature and HRT (Table 5). The biogas
productivity, but not the methane yield, was positively
affected by an increased solid loading. The effects of
hydraulic retention time and the solids loading can be
combined into what has been called the LR or organic
loading rate (OLR) which describes the amount of organic
material (expressed as VS) feed daily per volume of
working digester (g VS day1 l1 or kg VS day1 m3 Þ. The
LR depends directly on the % manure in the slurry and is
inversely proportional to the HRT. The CH4 yield
decreases with increasing LRs for both manures and at
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72 R. Alvarez et al. / Biomass and Bioenergy 30 (2006) 66–75

Table 5
Statistical analysis of the logarithmically transformed models

EFFECTS ANOVA (Partial sum of squares)

Term Effect SS Contribtn. DF F value Prob4F

Cow—methane yield
A 0.1352 0.0366 2.7907
B* 0.6748 0.9108 69.4941 1 33.39 0.0022
C 0.3629 0.2634 20.1010 1 9.66 0.0266
D 0.1391 0.0387 2.9535
AB 0.0793 0.0126 0.9603
AC 0.0633 0.0080 0.6114
AD 0.1423 0.0405 3.0890
Residual 0.1400 5
Cor. total 1.3100 7

Cow—biogas production
A 0.1006 0.0203 2.1721 1 4.15 0.1345
B 0.5976 0.7143 76.6114 1 146.27 0.0012
C 0.0823 0.0135 1.4532
D 0.2530 0.1281 13.7363 1 26.23 0.0144
AB 0.0219 0.0010 0.1028
AC 0.0085 0.0001 0.0153
AD 0.1659 0.0551 5.9090 1 11.28 0.0438
Residual 0.0150 3
Cor. total 0.9300 7

Llama—methane yield
A 0.0842 0.0142 0.8271
B 0.7984 1.2750 74.3015 1 39.79 0.0015
C 0.3747 0.2807 16.3607 1 8.76 0.0315
D 0.1515 0.0459 2.6750
AB 0.1051 0.0221 1.2866
AC 0.0533 0.0057 0.3311
AD 0.1902 0.0724 4.2180
Residual 0.1600 5
Cor. total 1.7200 7

Llama—biogas production
A 0.0470 0.0044 0.3551
B 0.7079 1.0021 80.5002 1 44.67 0.0011
C 0.0902 0.0163 1.3081
D 0.2555 0.1306 10.4884 1 5.87 0.0607
AB 0.1991 0.0792 6.3655
AC 0.0647 0.0084 0.6732
AD 0.0439 0.0039 0.3094
Residual 0.1100 5
Cor. total 1.2400 7
*
Significant factor.

both temperatures studied (Fig. 3). This agrees with results
from other studies [4,25]. On the other hand with respect to
the biogas productivity the manure percentage was more
important than HRT (Table 5), and an almost linear
relationship between the methane productivity and the
substrate loading for cow manure was observed at 35 and
11
C (Figs. 4a). This is in accordance with previously
reported results limited to mesophilic conditions for certain
types of animal wastes (swine, beef, poultry, and dairy
manure). This linear relationship will hold only up to a
certain LR (depending on the type of waste) above which
biological stress factors become inhibitory to the produc-
tivity [2,26]. The behaviour of llama manure (Figs. 4b) is
similar to cow manure at low temperature and mesophilic
conditions until a LR of 2.22, after which there appears to
be a change of slope.
Somewhat surprisingly, the pressure effect in the range
studied (495–760 mmHg) was not significant. Although, the
effect could possibly be found significant in an extended
study, the effect is nevertheless small, and low pressure will
not negatively affect the biogasification of the manures in a
small scale distributed process on the Altiplano.
 ARTICLE IN PRESS
CH4yield (I CH4kg−1VSadded) R. Alvarez et al. / Biomass and Bioenergy 30 (2006) 66–75 73

150 can be calculated from Bushwell’s formula [27]. Moller

et al. [28] found that due to the higher proportion of lipid
120 in pig manure, the Bu was higher in pig ð516 
90 11 l kg1 VSÞ manure than in cattle manure ð468
6 l kg1 VSÞ. The experimentally obtainable biological
60 methane potential from VS added ðl CH4 kg1 VSÞ is
30 referred to as the ultimate methane yield (Bo). The
quantity, Bo, represents the maximum quantity of methane
0 that can be produced per mass of VS load in anaerobic
0 1 2 3 4 5
digestion process in which the residence time approaches
(a) Loading Rate (g VS I-1d-1)
infinity. There is significant spread in the estimates of Bo
from several studies [21,28,29], with values for cattle
CH4yield (I CH4kg−1VSadded)

150
manure ranging from 0.13 to 0:285 m3 kg1 VS, and for
120 swine manure between 0.27 and 0:36 m3 kg1 VS. This is
likely a result of the dependence on breed and growth stage
90
of the animal, diet, which vary significantly from farm to
60 farm but perhaps more importantly from country to
country [28,29]. In the present study, the methane yield
30
obtained with cow manure at 11
C was between 6.40 and
0 33:57 l CH4 kg1 VS, whereas at 35
C the methane yield
0 1 2 3 4 5 was between 49.57 and 131:33 l CH4 kg1 VS. The low
-1day-1)
(b) Loading Rate (g VS I yields obtained could be attributed to the quality of
the VS in the manure, and in particular the low nitrogen
Fig. 3. Methane yield as function of loading rate from (a) cow manure
and (b) llama manure at 35
C ð’Þ, and 11
C ðEÞ. content. This indicates a relative low content of proteins
and lipids and higher content of material more difficult to
digest, most likely a consequence of the feed source
0.3 of the cows in the Bolivian highland. The methane
CH4 Productivity (l l−1d−1)

yield from llama manure is somewhat lower than for cow

manure, 35.56–84:08 l CH4 kg1 VS at 35
C and 3.28–
0.2 19:30 l CH4 kg1 VS at 11
C. On the other hand, the
biogas productivity (Fig. 4) shows that the llama manure
is somewhat higher than for cow manure at 35
C and
0.1
somewhat lower at 11
C. Consequently, as high a mean
temperature as possibly is desired for digestion of the llama
0.0 manure. The practical challenge is to avoid the temperature
0 1 2 3 4 5 fall during the night (e.g. by a suitable insulation systems)
(a) Loading Rate (g VS l−1d−1) and to take advantage of the heat of the sun during the
day. A retention time of 50 days seems appropriate. The
0.3
results indicate that llama manure is in fact a good source
CH4 Productivity (l l−1d−1)

for locally distributed biogas production on the Altiplano.

0.2 Because of the high solids content of the llama manure
(Table 2), the energy requirements of a housing could be
satisfied with a relatively low amount of collected llama
0.1 manure given its energy density. The methane yield of
84:08 l CH4 =kg VS load from llama manure contains
0.0
1:46 kJ g1 manure ð0:4 k Wh kg1 Þ as compared to
0 1 2 3 4 5 0:66 kJ g1 ð0:18 kWh kg1 Þ for cow manure (methane yield
(b) Loading Rate (g VSl−1d−1) of 131:31 CH4 kg1 VS added).

Fig. 4. Methane productivity as function of loading rate from (a) cow 5. Conclusions
manure and (b) llama manure at 35
C ð’Þ, and 11
C ðEÞ.

The results of this study show that technically it is

4.3. Biogas and methane productivity
The maximum theoretical amount of methane produced
per amount of VS consumed ðl CH4 kg1 VSÞ, denoted Bu,
feasible to produce biogas from anaerobic digestion of
llama and cow manure from the Andean Altiplano also at
low pressure. The methane yield and the daily biogas
production is highly dependent on the temperature, but the
HRT and manure content in the feed are also significant
 ARTICLE IN PRESS
74 R. Alvarez et al. / Biomass and Bioenergy 30 (2006) 66–75

Table 6
Characteristics of fresh manures from different animals

Parameter Unitsa Cowb Llamab Dairyc Beefc Vealc Swinec Sheepc Goatc Horsec Broilerc Turkeyc Duckc

Total solids % 15.58 59.53 12 8.5 5.2 11 11 13 15 22 12 31

Volatile solids % 12.9 44.31 10 7.2 2.3 8.5 9.2 — 10 17 9.1 19
pH 7.8 7.8 7 7 8.1 7.5 — — — — — —
Total Kjeldahl % 0.19 1.13 0.45 0.34 0.27 0.52 0.42 0.45 0.3 1.1 0.62 1.5
nitrogen
Ammonia % 0.008 0.083 0.079 0.086 0.12 0.29 — — — — 0.08 —
nitrogen
Total % 0.037 0.31 0.094 0.092 0.066 0.18 0.087 0.11 0.071 0.3 0.23 0.54
phosphorus
Potassium % — — 0.29 0.21 0.28 0.29 0.32 0.31 0.25 0.4 0.24 0.71
Iron g/kg 0.607 1.429 0.12 0.078 0.003 0.16 0.081 — 0.16 — 0.75 —
Oil and greasy % 12.66 21.85 — — — — — — — — — —
a
All values based on wet weight.
b
Calculated from Table 1.
c
Adapted from ASAE D384.1 FEB03 [27].

factors, although to a lesser extent than the temperature.
The low pressure (495 mmHg, mean atmospheric pressure
of the Bolivian highlands) does not significantly affect the
biogas production. Of the materials studied, llama manure
appears to be the best raw material. In fact, the llama
manure contains more volatile solid, nitrogen, and
phosphorous than previously has been reported for a
number of other manures, e.g. cow, swine, sheep, goat,
horse, broiler, and turkey manure, and it is as easy to digest
as cow manure.
Acknowledgements
This work was financially supported by SIDA, the
Swedish International Development Cooperation Agency.
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