Waste Management 48 (2016) 199–208

Contents lists available at ScienceDirect

Waste Management
journal homepage: www.elsevier.com/locate/wasman

Comparison of start-up strategies and process performance during semi-

continuous anaerobic digestion of sugarcane filter cake co-digested with
bagasse
Leandro Janke a,b,⇑, Athaydes F. Leite c, Marcell Nikolausz c, Claudemir M. Radetski d, Michael Nelles a,b,
Walter Stinner a
a
Department of Biochemical Conversion, Deutsches Biomasseforschungszentrum gemeinnützige GmbH, Torgauer Straße 116, 04347 Leipzig, Germany
b
Faculty of Agricultural and Environmental Sciences, Department of Waste Management, University of Rostock, Justus-von-Liebig-Weg 6, 18059 Rostock, Germany
c
Department of Environmental Microbiology, Helmholtz Centre for Environmental Research – UFZ, Permoserstraße 15, 04318 Leipzig, Germany
d
Laboratório de Remediação Ambiental, Universidade do Vale do Itajaí, Rua Uruguai 458, 88302-202 Itajaí, Brazil

a r t i c l e i n f o a b s t r a c t

Article history: The anaerobic digestion of sugarcane filter cake and the option of co-digestion with bagasse were inves-
Received 26 April 2015 tigated in a semi-continuous feeding regime to assess the main parameters used for large-scale process
Revised 3 November 2015 designing. Moreover, fresh cattle manure was considered as alternative inoculum for the start-up of bio-
Accepted 3 November 2015
gas reactors in cases where digestate from a biogas plant would not be available in remote rural areas.
Available online 12 November 2015
Experiments were carried out in 6 lab-scale semi-continuous stirred-tank reactors at mesophilic condi-
tions (38 ± 1 °C) while the main anaerobic digestion process parameters monitored. Fresh cattle manure
Keywords:
demonstrated to be appropriate for the start-up process. However, an acclimation period was required
Sugarcane waste
Anaerobic digestion
due to the high initial volatile fatty acids concentration (8.5 g L
1). Regardless the mono-digestion of filter
Start-up cake presented 50% higher biogas yield (480 mL gVS
1) than co-digestion with bagasse (320 mL gVS
1)
Biogas process during steady state conditions. A large-scale co-digestion system would produce 58% more biogas
Reactor performance (1008 m3 h
1) than mono-digestion of filter cake (634 m3 h
1) due to its higher biomass availability for
biogas conversion. Considering that the biogas production rate was the technical parameter that dis-
played the most relevant differences between the analyzed substrate options (0.99–1.45 m3 biogas m3
d
1). The decision of which substrate option should be implemented in practice would be mainly driven
by the available construction techniques, since economically efficient tanks could compensate the lower
biogas production rate of co-digestion option.
Ó 2015 Elsevier Ltd. All rights reserved.

1. Introduction waste, which in most cases are still not being properly managed
from the energy point of view (De Carvalho Macedo, 2007). In this
The production of sugar and ethanol based on sugarcane as context, bagasse, a solid waste derived from the extraction of sug-
feedstock is responsible for generation of different types of organic arcane juice, is generated in large amounts (260 kg per ton of cane),
and usually used as fuel in low-efficiency cogeneration systems or
sold by the sugarcane plants to another end-uses (e.g. animal feed-
Abbreviations: AD, anaerobic digestion; C:N, carbon to nitrogen; C:P, carbon to ing) (Bressan Filho, 2011; Nogueira et al., 2008). On the other hand,
phosphorus; C:S, carbon to sulfur; FCM, fresh cattle manure; FM, fresh matter; HRT,
filter cake, produced during the clarification (physical–chemical
hydraulic retention time; MIX, mixture of digestates; NFC, non-fiber carbohydrates;
NH4-N, ammonium-nitrogen; OLR, organic loading rate; SBP, specific biogas process) of the sugarcane juice, is generated in lower amounts than
production; SCSTR, semi-continuous stirred-tank reactor; SUC, specific upgrading bagasse (35–40 kg per ton of cane), however it is mostly applied as
cost; TBP, theoretical gas potential; TS, total solids; VFA, volatile fatty acids; VOA, organic fertilizer on the sugarcane fields without any previous
volatile organic acids; VOA/TIC, ratio of volatile organic acids and total inorganic energy recovery (Janke et al., 2015a).
carbonate to calcium carbonate; VS, volatile solids.
⇑ Corresponding author at: Department of Biochemical Conversion, Deutsches Anaerobic digestion (AD) is a promising strategy to manage
Biomasseforschungszentrum gemeinnützige GmbH, Torgauer Straße 116, 04347 such type of waste, since as a result of the biochemical process
Leipzig, Germany. in which complex organic matter is degraded to CH4 and CO2 by
E-mail address: [email protected] (L. Janke).

http://dx.doi.org/10.1016/j.wasman.2015.11.007
0956-053X/Ó 2015 Elsevier Ltd. All rights reserved.
200 L. Janke et al. / Waste Management 48 (2016) 199–208

various types of microorganisms biogas could be produced and 2. Materials and methods
used as fuel to improve the energy balance of the sugarcane plants
(Janke et al., 2015b; Leite et al., 2015a). However, several factors 2.1. Substrates and inocula
such as temperature, pH, organic loading rate (OLR), hydraulic
retention time (HRT), balance of nutrients and presence of inhibi- Samples of sugarcane filter cake and bagasse were collected
tors must be considered for an efficient AD process. Furthermore, from a distillery plant in the State of Goiás (Brazil) during the
the microbial community and the quality of the inoculum used 2012/2013 season, transported to Germany in sealed plastic bags
for the start-up of an anaerobic reactor are also considered decisive and kept under low temperature (i.e. 4 °C) until its use. A large-
factors for a successful biogas production (Cho et al., 2013; Moset scale biogas plant that uses maize silage and fresh cattle manure
et al., 2014). (FCM) as substrate provided FCM that was used for the start-up
In previous studies (Janke et al., 2014; Leite et al., 2015b) our of two semi-continuous reactors. A mixture of several digestates
group has already assessed the possibility of using these type of (hereafter referred as MIX) from mesophilic lab-scale reactors were
waste for biogas production in batch tests. However, the feed used for the start-up of four other semi-continuous reactors. To
regime and high proportion of inoculum used during batch tests avoid inlet and outlet pipes from clogging, both inocula were
do not allow an adequately assessment of anaerobic reactors sieved prior to inoculation in the reactors. Tap water was utilized
start-up, neither possible process inhibition during digestion of to keep the total solids of the feed below 15% for the wet fermen-
the substrates. Therefore, understanding the reactor’s behavior tation process.
during the start-up phase can be attained only by using a similar
feeding regime applied in large-scale applications (semi- 2.2. Semi-continuous feeding experiments
continuous system).
For large-scale applications, it is well known that using diges- Six lab-scale semi-continuous stirred-tank reactors (SCSTR)
tate taken from a stable working digester could be a good strategy with 5 L total volume and 3 L working volume were carried out
to overcome the start-up challenges (Kobayashi et al., 2009). How- in these experiments. The reactors were continuously stirred
ever, in countries where biogas technology is not established in the (100 rpm) using a central stirrer with helix shaped blades located
market yet, plant operators must find alternative sources of inocu- in the lower part of the reactors. The operation temperature was
lum suitable for the start-up period of an anaerobic reactor. In this kept under mesophilic conditions (38 ± 1 °C) by recirculating hot
case, the utilization of animal waste, such as cattle manure, could water through the double-walled reactors.
be a useful strategy, since such material is rich in microorganisms Each of the three following experiments performed in our study
from animal digesting system, as well important macronutrients was carried out in duplicate with the same feeding regime (once
and trace elements (Seadi et al., 2008). per day). Reactors R3.3 and R3.4 were fed with filter cake and
According to earlier studies (Kayhanian and Rich, 1995; MIX for start-up. Reactors R3.5 and R3.6 were fed in a co-
Mccarty, 1964), another factor that can influence the performance digestion system with filter cake (70%) and bagasse (30%) on fresh
of the AD process is the nutrient content of the substrates. If a cer- matter basis, also using MIX for start-up. Reactors R3.7 and R3.8
tain substrate has too high C:N ratio and consequently nitrogen were fed with the same co-digestion proportion, however using
deficiency, it may negatively affect the functioning of the microbial FCM as inoculum. Detailed information about the different feeding
community. Thus, a direct effect on their ability to produce rates, OLR and HRT are listed in Table 1.
enzymes that are needed for the carbon utilization, causing an
incomplete conversion of the substrates, resulting in lower CH4 2.3. Analytical methods
yields. On the other hand, substrates that contain high levels of
nitrogen can cause inhibition to the AD process via accumulation For all samples, total solids (TS) and volatile solids (VS) were
of toxic ammonia (NH3) produced from protein degradation or by analyzed according to VDI 4630 (2006). Nutritional values were
urea conversion (Lv et al., 2014). determined according to Weender followed by Van Soest methods.
The characteristics of different sugarcane waste assessed by By the Weender method raw protein, raw fat, non-fiber carbohy-
Leite et al. (2015b), showed that bagasse has a C:N variation of drates (NFC) and raw fiber are determined. Van Soest method
90–101:1 along an operating season, which is higher than the allows the determination of the remaining carbohydrates and lig-
range of values (20–40:1) recommended by others (FNR, 2012). nin fractions from the neutral detergent fiber (NDF), which repre-
Meanwhile, previous studies (López González et al., 2013) showed sents hemicellulose, cellulose, lignin and ash, acid detergent fiber
that filter cake has a C:N ratio of 26:1, that is around the lowest (ADF) represented by cellulose, lignin and ash, and the lignin con-
recommended limit. tent depicted by the acid detergent lignin (ADL). Detailed descrip-
Considering that filter cake is a waste stream that currently is tion of the methods was previously published by Liebetrau et al.
not used for any energy purpose, it makes sense to use such type (2015). To determinate the major elements contained in each sug-
of biomass on the AD process to produce biogas. This would arcane waste, dried samples were pre-treated with a mixture of
enhance the energy balance of sugarcane plants without losing HNO3/H2O2/HF and latter neutralized with H3BO3, and the result-
the essential nutrients for the sugarcane cultivation. Additionally, ing clear solution was analyzed by inductively coupled plasma
bagasse that is the major solid waste produced on-site by the sug- atomic spectrometry (ICP-OES, ThermoFischer iCAP6200) accord-
arcane plants, could be an interesting co-substrate to balance the ing to standard procedures (DIN, 2011a, 2011b, 2002).
C:N of filter cake and improve energy production in the biogas The daily biogas production in each of the semi-continuous
system. Thus, the objectives of the present study were to (i) assess reactors was measured by a milligascounter type MGC-10 (Ritter,
cattle manure as alternative inoculum for the start-up phase of Bochum, Germany), and corrected to standard temperature and
semi-continuous anaerobic reactors; (ii) compare the process pressure conditions (273.15 K and 101.325 kPa), and the specific
performance during semi-continuous mono-digestion of filter cake biogas production (SBP) was presented in norm milliliters per g
versus the option of co-digestion with bagasse; and (iii) analyze of VS (mL gVS
1). The composition of the produced biogas (CH4,
both substrate options (mono-digestion and co-digestion) on the CO2 and O2) was measured twice a week in the headspace of the
main parameters used for the AD process design integrated to a reactors by using a GA2000 Landfill Gas Analyzer (Geotechnical
large-scale sugarcane plant. Instruments Ltda., UK).
 L. Janke et al. / Waste Management 48 (2016) 199–208 201

Table 1
Overview of the semi-continuous feeding experiment.

Reactors Inoculum C:N ratio Phase Period (day) Filter cake Bagasse Water HRT (days) OLR (gVS L d
1)
Input (g day
1) Input (g day
1) Input (mL day
1)
R3.3 MIX 24:1 Start-up 0–41 36.5 – 50 34.7 2.0
R3.4 42–69 45.7 – 50 31.4 2.5
Steady I 70–113 54.8 – 50 28.6 3.0
Steady II 114–137 54.8 – 75 23.1 3.0
R3.5 MIX 41:1 Start-up 0–41 15.78 6.78 50 41.4 2.0
R3.6 42–69 19.73 8.45 50 38.4 2.5
Steady I 70–113 23.67 10.15 50 35.8 3.0
Steady II 114–137 23.67 10.15 75 27.6 3.0
R3.7 FCM 41:1 Start-up 0–3 15.78 6.78 50 41.4 2.0
R3.8 4–20 7.89 3.39 25 82.7 1.0
21–41 15.78 6.78 50 41.4 2.0
42–69 19.73 8.45 50 38.4 2.5
Steady I 70–113 23.67 10.15 50 35.8 3.0
Steady II 114–137 23.67 10.15 75 27.6 3.0

Five days per week pH was measured immediately in fresh

Table 2
digestate of each reactor with a pH-electrode (WTW type pH Main characteristics of the sugarcane plant used as reference to assess the large-scale
3310 Sentix 41, Germany). Once a week fresh digestate samples biogas application.
were also centrifuged in 10,000 rpm during 10 min at 10 °C and
Characteristic Value Unit
supernatant liquid was used after filtration to subsequent analysis.
Filtered samples (10 mL) were used for the quantification of all Working days 232 days year
1
Sugarcane processed 2.0  106 TC year
1
volatile organic acids (VOA) and a ratio of total inorganic carbonate Specific filter cake generation 35 kgFM TC
1
to calcium carbonate (VOA/TIC, gVOA/gCACO3) measurement in a Total filter cake generation 70  103 tonFM year
1
Titration Excellence T90 titrator (Mettler-Toledo GmbH, Switzer- Specific bagasse generation 260 kgFM TC
1
land). The concentrations of the major volatile fatty acids (VFA), Total bagasse generation 520  103 tonFM year
1
including acetic- and propionic acid, were determined by gas chro- TC: tons of cane.
matography using a 5890 series II gas chromatograph (Hewlett
Packard, USA) equipped with a HS40 automatic headspace sampler
for large-scale process designing. Considering that for the Brazilian
(Perkin Elmer, USA) and an Agilent HP-FFAP column
conditions depending on the energy balance of the sugarcane
(30 m  0.32 mm  0.25 lm). Ammonium-nitrogen (NH4-N g L
1)
plants a surplus of bagasse is expected (Nogueira et al., 2008),
was determined from 500 lL filtered samples diluted with deion-
which corresponds to an average value of 8.5% of total bagasse gen-
ized water in a proportion of 1:1000 during the start-up phase
erated (Bressan Filho, 2011). The co-digestion proportion used in
and 1:500 during the steady phase with the Nessler method using
the laboratory experiments represents less than 6% of the total
a benchtop spectrophotometer (Hach-Lange DR 3900, Loveland,
bagasse generated by a sugarcane plant. Table 2 presents the main
US).
characteristics of the reference sugarcane plant used in this study.

2.4. Degradation index

3. Results and discussion
A degradation index was calculated to compare specific biogas
production achieved during semi-continuous feeding experiments 3.1. Substrate composition
to the theoretical biogas potential of the substrates, according to
Eq. (1). The main characteristics of substrates used during semi-
continuous feeding experiments are presented in Table 3. Filter
Dindex ¼ SBP=TBP ð1Þ cake, having an appearance similar to sludge, showed lower TS
where, and VS content once is derived from a physical–chemical treat-
Dindex: degradation index (fraction); ment process that removes soluble and insoluble impurities from
SBP: specific biogas production (mL gVS
1);
TBP: theoretical biogas potential (mL gVS
1); Table 3
Composition of sugarcane waste used during semi-continuous feeding experiments.

Theoretical biogas potential was calculated according to Eq. (2) Parameters Filter cake Bagasse Units
considering average values for carbohydrates, proteins and lipids TS 21.9 54.7 % FM
published by Weißbach (2009). VS 75.5 97.7 % TS
Raw protein 135.9 11.2 g kg
1 TS
TBP ¼ ðPTBP þ LTBP þ C TBP Þ ð2Þ Raw fat 37.7 8.2 g kg
1 TS
Carbohydrate NFC 115.5 80.8 g kg
1 TS
where, Cellulose 171.7 440.4 g kg
1 TS
PTBP: theoretical biogas potential of proteins (mL gVS
1); Hemi-cellulose 153.1 287.6 g kg
1 TS
LTBP: theoretical biogas potential of lipids (mL gVS
1); Lignin 141.4 149.4 g kg
1 TS
Raw ash 244.6 22.4 g kg
1 TS
CTBP: theoretical biogas potential of carbohydrates (mL gVS
1);
Carbon 40.8 49.7 % TS
Nitrogen 1.72 0.55 % TS
2.5. Preliminary assessment for large-scale process designing Phosphorus 0.6 0.01 % TS
Sulfur 0.7 0.02 % TS
Total Kjeldahl nitrogen 21.8 1.82 g kg
1 TS
Results from semi-continuous digestion were used to assess the
C:N ratio 24:1 90:1 –
effects of different substrate options on the main parameters used
202 L. Janke et al. / Waste Management 48 (2016) 199–208

the sugarcane juice. Bagasse presented higher carbohydrate con- digestion, which in turn represents 50% lower than the theoretical
tent, especially in form of cellulose and hemicellulose, since it is biogas potential calculated based on substrate nutritional values.
basically composed by sugarcane fibers. In this way, although both During the unstable period in the process, the co-digestion reactors
substrates displayed similar lignin content, it is expected that the behaved similarly to the mono-digestion, decreasing the SBP below
negative effects of lignin during AD would be more significant in average as result of the VFA accumulation and increasing above
bagasse than filter cake because bagasse has 124% higher carbohy- average afterward due to the same reasons previously described.
drates in fibrous form. Additionally, the CH4 content found in biogas from the co-
Both substrates presented large differences in terms of digestion was lower than in mono-digestion during stable condi-
macronutrients. The carbon to nitrogen ratio of filter cake (24:1) tions and lower as well during the unstable period of the process.
is within the optimum range for AD (20–40:1), while bagasse pre- This can be explained by different substrate compositions, since
sented much lower nitrogen content, resulting in higher C:N ratio according to Weißbach (2009) carbohydrate-based substrates
of 90:1. If phosphorus and sulfur are also considered, the unbalance would produce around 50–55% (co-digestion of filter cake and
of nutrients is even more evident. Filter cake presented relatively bagasse) of CH4 in biogas, while a share of protein could increase
low C:P and C:S ratios (68:1 and 58:1), what could lead in extreme CH4 content up to 60–65% (mono-digestion of filter cake).
cases to competition of sulfate-reduction with methanogenesis, Furthermore, the higher cellulose and hemicellulose content
reducing the conversion of organic acids into biogas, and also neg- found in bagasse might have been another factor that has influ-
atively influencing the bioavailability of trace elements inside the enced the differences in SBP between the mono-digestion and
bioreactors (Gustavsson et al., 2013; Schmidt et al., 2014a). co-digestion with bagasse. According to Montgomery and
Meanwhile, bagasse presented much higher values for C:P Bochmann (2014) the recalcitrant presence of lignin may have
(4970:1) and C:S 2485:1, supporting the idea of a co-digestion hampered the hydrolysis of such fibrous materials resulting in a
strategy to balance those nutrients in order to improve the degrad- lower conversion of the volatile solids into biogas.
ability and energy production from both substrates.
3.2.2. Fresh cattle manure (FCM)
3.2. Start-up during the semi-continuous experiment A completely different situation was found during the start-up
of co-digestion reactors inoculated with FCM (reactors R3.7–3.8).
3.2.1. Mixture of digestates (MIX) Due to the high VFA concentration in the inoculum, composed by
All reactors that used MIX as inoculum performed a relatively 60% of acetic acid, 20% of propionic acid and 10% of butanoic acid,
stable process during the start-up phase (days 0–69), except in a among other minor acids, both reactors were subjected to a greater
short period between days 20–30 for co-digestion with bagasse process instability demanding a temporary feed reduction (OLR at
(reactors R3.5–3.6) and days 30–40 for mono-digestion of filter 1.0 gVS L d
1) during days 4–20, until VFA levels stabilized and
cake (reactors R3.3–3.4). In both cases acetic acid was the predom- VOA/TIC returned to the safe ratio of <0.5.
inant accumulated VFA (around 90% of total VFA) suggesting a Therefore, the biogas production monitored during this period
temporarily failure of the acetoclastic methanogenesis pathway cannot be considered realistic from the substrate potential because
during the adaptation of the microbial community to the substrate most of the performed SBP were derived from the inoculum VFA
(Banks et al., 2012). conversion into biogas, which after further observation showed a
However, the same amount of accumulated acids caused differ- drastic SBP reduction immediately after the consumption of VFA
ent negative effects when both substrate options are compared. originated from the inoculum (Fig. 3).
The VOA/TIC increased higher in mono-digestion than in co- Despite of the high VFA concentration, the measured pH during
digestion, since different concentrations of NH4-N, at the moment the entire start-up phase was in an optimum range considered for
when VFA started to accumulate, were found (Figs. 1 and 2). The AD, mainly due to the high buffer capacity provided by FCM, of
higher NH4-N concentration found during process instability of which the initial concentration of NH4-N was around 117% higher
co-digestion (0.80 g L
1) could have helped to buffer the system than found in the other co-digestion system inoculated with MIX.
against the more severe pH drop found during the process instabil- On one hand the higher NH4-N concentration provided by FCM
ity of mono-digestion (NH4-N of 0.57 g L
1). Moreover, the process can be considered an advantage over the other evaluated inocu-
of inoculum washing-out that usually occurs immediately after the lum, since in case of process instability, the reactor inoculated with
start-up of SCSTRs seems to be the reason why the concentration of FCM could withstand higher unbalances caused by VFA accumula-
NH4-N was higher in co-digestion (instability at 20–30 days) than tion without major changes in the pH, which usually can happen
in mono-digestion (instability at 30–40 days) because the trend during start-up of anaerobic reactors due to process instabilities
of gradually NH4-N depletion can be seen in all reactors during caused by OLR increase and slow adaptations of microorganisms
the entire experiment period. to the substrate.
Regarding biogas production, the mono-digestion showed a 41% On the other hand, the high VFA concentration found in FCM
lower specific biogas production (SBP) than the theoretical biogas would demand a proper acclimation of inoculum to the reactor envi-
potential calculated based on substrate nutritional values (see Eq. ronment to reduce the VFA and VOA/TIC ratio to a safe level, which
(2)). In general, the SBP did not vary considerably during the entire could delay the normal start-up of a large-scale biogas plant by at
phase, except during the process instability occurrence described least 20 days. On the contrary, if the acclimation period is not
previously. During this short period, a lower than average SBP respected, the VFA concentration could reach a certain level where
was observed for both mono-digestion reactors, caused by VFA the NH4-N would not be able to keep an optimum pH value for
accumulation, followed by a rapidly increase over than the average methanogenesis (>6.5), leading to a possible failure of the AD process.
SBP. This can be explained by the normal substrate feeding without To compare the SBP of two co-digestion systems, it is appropri-
any interruption or reduction in OLR, which has provided more ate to consider values achieved after the acclimation period
VFA to biogas conversion at the moment when reactors recovered because until day 20 most part of SBP in reactors R3.7–3.8 was
normal performance. The CH4 content found in biogas was around derived from VFA degradation of the inoculum. In this way, the
60% (v/v) during stable conditions and 55% (v/v) during the short co-digestion system that used FCM as inoculum produced a similar
unstable process period (Table 4). SBP than observed in reactors inoculated with MIX. In contrast, CH4
When those results are compared to the co-digestion with in reactors R3.7–3.8 has presented a slightly higher concentration
bagasse, the SBP was 12.8% lower than in case of the mono- in comparison to reactors R3.5–R3.6.
 L. Janke et al. / Waste Management 48 (2016) 199–208 203

Fig. 1. Results during semi-continuous feeding experiment of filter cake using mix of inocula (Reactors R3.3 and R3.4). (A) Specific biogas production (SBP); (B) total volatile
fatty acids (VFA); (C) ratio of volatile organic acids with total inorganic carbonate to calcium carbonate (VOA/TIC); (D) pH; (E) Ammonium-nitrogen concentration (NH4-N).
204 L. Janke et al. / Waste Management 48 (2016) 199–208

Fig. 2. Results during semi-continuous feeding experiment of filter cake and bagasse using mix of inocula (Reactors R3.5 and R3.6). (A) Specific biogas production (SBP); (B)
total volatile fatty acids (VFA); (C) ratio of volatile organic acids with total inorganic carbonate to calcium carbonate (VOA/TIC); (D) pH; (E) Ammonium-nitrogen
concentration (NH4-N).
 L. Janke et al. / Waste Management 48 (2016) 199–208 205

Table 4
Specific biogas production during start-up and steady condition.

Reactors Phase SBP (mL gVS
1) CH4 (% v/v) TBP (mL gVS
1) Dindex
R3.3–R3.4 Start-up 382 60.0 0.59
Steady I 480 57.9 650 0.74
Steady II 471 58.7 0.72
R3.5–R3.6 Start-up 333 55.0 0.50
Steady I 320 53.0 0.48
Steady II 277 54.8 0.41
671
R3.7–R3.8 Start-up 326 54.4 0.49
Steady I 320 52.3 0.48
Steady II 277 55.6 0.41

SBP: specific biogas production.

TBP: theoretical biogas potential.
Dindex: degradability index.

These findings are considered unexpected since the higher was kept in 35.8 days, and reduced by 13.2% when the HRT
availability of nitrogen in the form of NH4-N provided by FCM decreased to 27.6 days. However, as opposed to the mono-
should have increased the conversion of the co-digestion substrate digestion, in both co-digestion systems the SBP observed during
(C:N of 41:1) into biogas. This means that the type of inoculum has the steady phase was not higher than found during the reactors
not influenced significantly the final energy production for the co- start-up (excluding the days 4–20 in reactors R3.7–R3.8), even con-
digestion with bagasse, leading to the conclusion that utilization of sidering an average increase in the biomass inside the reactors TS
FCM did not result in any advantage over MIX, at least from the from 4% to 8%. In this case, the higher dependency of additional
energy point of view. nutrients necessary for the co-digestion with bagasse (poorer in
macronutrients) was possible supplied by both inocula during
3.3. Steady condition during the semi-continuous experiment the reactor’s start-up.
Indeed, the substrates composition played a major role during
3.3.1. Mono-digestion of filter cake the semi-continuous experiments. The additional carbon supplied
After the start-up phase and inoculum wash-out, the OLR of all by bagasse in the co-digestion system was not able to improve
reactors was increased to 3.0 gVS L d
1 at day 70 and stayed until the overall AD performance, mainly due to the form in which car-
the end of the experiment at day 137. During this period the mono- bon was provided (fibrous carbohydrates). Moreover, the low con-
digestion of filter cake (reactors R3.3–3.4) did not present any sign centration of phosphorus and sulfur found in bagasse has also
of process unbalance, even when the NH4-N and pH achieved their negatively influenced the biogas production, since both elements
lowest values observed during the entire experiment. are important during microbial growth (Kayhanian and Rich,
The mono-digestion system achieved a SBP of 480 mL gVS
1 1995). In this case, a supplementation strategy based on phosphate
(Dindex of 0.74) during the period when the HRT was kept in and sulfate are recommended (Scherer et al., 2009), however
28.6 days (70–113 days), and slightly reduced when the HRT was demanding special attention to avoid excessive costs for the anaer-
changed to 23.1 days during the days 114–137. Such higher biogas obic system.
production found during steady phase (+25%) in comparison to the
start-up phase is explained by a higher biomass concentration
inside the reactors found at day 137 (TS of 10.6% and VS 6.07%) 3.4. Preliminary assessment for large-scale process designing
in comparison to the biomass found during the beginning of exper-
iment (TS of 4.02% and VS of 1.42%). In order to analyze the positive and negative aspects of different
substrate options, an assessment of the main parameters used for
3.3.2. Co-digestion with bagasse designing the AD process integrated to a sugarcane plant with
In contrast to the results obtained during the mono-digestion of the capacity to process 2 million tons of cane per year is presented
filter cake, all reactors used for the co-digestion with bagasse in Table 5.
(R.3.5–3.8) presented an acid accumulation immediately after the Considering the same substrate proportions used during our
OLR increase from 2.5 to 3.0 gVS L d
1 at day 70 until day 105, this semi-continuous digestion experiments, the co-digestion option
time mainly composed of propionic acid (90% of total VFA). Accord- would have 27,606 tons of VS y
1 to be used as substrate for biogas
ing to previous studies conducted by Schmidt et al. (2014b), propi- conversion, while mono-digestion would have 11,574 tons of VS
onate oxidizing bacteria are sensitive to the lack of trace elements, y
1. Such a difference is not only explained by higher substrate uti-
especially iron and nickel depletion, which could be the reason of lization during co-digestion (35 kg of filter cake per ton of cane +
such VFA accumulation, since at this time of the experiment most 15 kg of bagasse per ton of cane), but also by higher TS and VS con-
part of the macronutrients and trace elements contained in MIX tent found in bagasse in comparison to filter cake.
and FCM were already washed-out from the reactors. Therefore, even considering that mono-digestion presented
Moreover, due to the lower buffer capacity as a result of the higher SBP during continuous digestion, the hourly biogas produc-
decreasing NH4-N levels in comparison to the start-up phase, the tion of co-digestion (1008 m3 h
1) would be 58% higher than
VOA/TIC ratio achieved its highest value during the entire experi- mono-digestion (634 m3 h
1).
ment (0.57). As a consequence, the pH dropped to its lowest However, such higher biogas production during co-digestion
observed level (6.45), which could have caused an inhibition of could only be achieved if the HRT would be 35.8 days, which is
methanogenic activity leading to complete process failure. 25% longer than the HRT needed for mono-digestion (28.6 days).
In the meantime, the SBP of the co-digestion system (biogas Thus, when the sizes of the reactors are calculated, the differences
production was measured only in reactors R3.5, R3.7 and R3.8 between the substrate options are even larger, and co-digestion
due to a technical failure of the milligascounter in reactor R3.6) would need a 24,303 m3 reactor, while mono-digestion a 10,499
reached the value of 320 mL gVS
1 (Dindex of 0.48) when the HRT m3 reactor. Nevertheless, differences on biogas production rate
206 L. Janke et al. / Waste Management 48 (2016) 199–208

Fig. 3. Results during semi-continuous feeding experiment of filter cake and bagasse using fresh cattle manure as inoculum (Reactors R3.7 and R3.8). (A) Specific biogas
production (SBP); (B) total volatile fatty acids (VFA); (C) ratio of volatile organic acids with total inorganic carbonate to calcium carbonate (VOA/TIC); (D) pH; (E) Ammonium-
nitrogen concentration (NH4-N).
 L. Janke et al. / Waste Management 48 (2016) 199–208 207

Table 5 with bagasse was 31% lower than the mono-digestion of filter
Main parameters used for designing the AD process in large-scale. cake.
Substrates Filter Bagasse Co- Unit
cake digestiona
Acknowledgements
Fresh matter 70,000 30,000 100,000 ton y
1
TS 15,330 16,410 31,740 ton y
1
The authors would like to acknowledge the support of the
VS 11,574 16,032 27,606 ton y
1
SBP 480 – 320 mL gVS
1 Brazilian National Scientific Counsel (CNPq) under the Program
CH4 content 60 – 55 % v/v Science without Borders for the financial support of the PhD stu-
OLR 3.0 – 3.0 gVS L d
1 dents Leandro Janke (237938/2012-0) and Athaydes Leite
HRTb 28.6 – 35.8 days (202024/2012-1). The present research was partially financed by
Biogas production 634 – 1008 m3 h
1
the i-NOPA Project ‘‘Sustainable bioeconomy in Brazil: Bioenergy
Size of reactor 10,499 – 24,303 m3
Biogas production 1.45 – 0.99 m3 biogas m3 from biogas using various types of waste substrates from the
rate d
1 Brazilian bioethanol industry”.
SUCc 1.60 – 1.34 EUR ct kW h
1
a
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