Environmental Technology & Innovation 10 (2018) 215–222

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Environmental Technology & Innovation

journal homepage: www.elsevier.com/locate/eti

Biodegradation of rice straw under anaerobic digestion

Cristian Candia-García, Liliana Delgadillo-Mirquez *, Mauricio Hernandez
Faculty of Engineering, University of Ibague, Colombia

highlights

• Different varieties of rice straw in the state of Tolima–Colombia does not present significant differences between compositional values.
• High solid loading rates inhibit biogas production during anaerobic digestion period.
• Highest production of biogas was reached with inoculum to substrate ratio (I/S) = 0.8 with a biogas yield from 410 L/kgVS.
• Cellulose structure showed the highest efficiency degradation (50.10%) in comparison with hemicellulose and Lignin structures (44.17%
and 32.62%, respectively).

article info a b s t r a c t
Article history: To mitigate environmental impacts caused by illegal burning of rice straw in rural fields,
Received 23 June 2017 anaerobic digestion of rice straw could be an excellent alternative for the treatment of
Received in revised form 13 February 2018 this sub-product of paddy rice harvest. The effect of inoculum/substrate ratio (I/S) on
Accepted 21 February 2018
biogas yield and biodegradation of rice straw was carried out in batch reactor under room
Available online 3 March 2018
temperature (25◦ to 27 ◦ C) and for 60 days of digestion. Results demonstrate a high biogas
Keywords: production (410 L/kgVS) at 0.8 of I/S ratio, with methane content over 70%. The modified
Lignocellulosic material Gompertz model was utilized to simulate the cumulative biogas yield from different I/S
Rumen content ratios. Degradation of volatile solids (VS), cellulose, hemicellulose, lignin, silica and cellular
Degradation efficiency content indicated that rumen content is an efficient degrader for lignocellulosic material.
Inoculum to substrate ratio This study provides useful parameters to evaluate the biogas production via anaerobic
Biogas yield digestion of rice straw, as a promising alternative energy in rural areas.
Co-digestion © 2018 Elsevier B.V. All rights reserved.

1. Introduction

Rice crops in Colombia reached an area of 450,000 ha have per year, distributed in 215 municipalities along the country
and produce over 2.4 million tons of paddy rice. This agricultural activity generates about 2% of Colombian GDP and it
is the means of economic survival for about 500,000 families, represented by 21,800 rice farmers. Overall, these farmers
have maintained traditional rice crop practices that represent high production costs and a very disadvantaged situation, in
comparison with rice imports from countries such the United States (Cardona, 2015). Besides, particular attention should
be paid to large quantities of rice straw produced in regular rice crops. According Universidad Nacional de Colombia, 1.4
ton rice straw is produced for each ton of paddy rice harvest. This lignocellulosic residue is often dumped or burned in an
open environment, which is not a recommended practice, in terms of environmental and ecological aspects of sustainable
development (Agencia de Noticias UN, 2014).
Rice straw is a lignocellulosic material, mainly composed by cellulose (34.89% ± 0.36% TS), hemicellulose (29.04% ± 0.54%
TS) and lignin (6.88% ± 0.76% TS). Previous studies have shown that these recalcitrant structures can be degraded by

* Corresponding author.
E-mail address: liliana.delgadillo@unibague.edu.co (L. Delgadillo-Mirquez).

https://doi.org/10.1016/j.eti.2018.02.009
2352-1864/© 2018 Elsevier B.V. All rights reserved.
216 C. Candia-García et al. / Environmental Technology & Innovation 10 (2018) 215–222

Nomenclature

Cellular C.de Degradation efficiency of Cellular content (%)

Cellular C.re Cellular content of digested residues of experimental group (%)
Cellular C.rs Initial Cellular Content of rice straw (%)
Cellulosede Degradation efficiency of Cellulose (%)
Cellulosere Cellulose content of digested residues of experimental group (%)
Cellulosers Initial Cellulose content of rice straw (%)
F733 Rice straw variety Fedearroz 768 from Colombia
F767 Rice straw variety Fedearroz 767 from Colombia
F768 Rice straw variety Fedearroz 768 from Colombia
F2000 Rice straw variety Fedearroz 2000 from Colombia
FER First experimental run
Hcellulosede Degradation efficiency of Hemicellulose (%)
Hcellulosere Hemicellulose content of digested residues of experimental group (%)
Hcellulosers Initial Hemicellulose content of rice straw (%)
I/S Inoculum to substrate ratio (VS/VS)
Ligninde Degradation efficiency of Lignin (%)
Ligninre Lignin content of digested residues of experimental group (%)
Ligninrs Initial Lignin content of rice straw (%)
SER Second experimental run
SLR Solid loading rate, kg/m3
Silicade Degradation efficiency of Silica (%)
T Anaerobic digestion time (d)
TSb TS of blank reactors (%)
TSde Degradation efficiency of TS (%)
TSre TS of digested residues of experimental group (%)
TSrs Initial TS content of rice straw (%)
Vw Working volume, mL
VSb VS of blank reactors (%)
VSde Degradation efficiency of VS (%)
VSre VS of digested residues of experimental group (%)
VSrs Initial VS content of rice straw (%)
Wb Weight of digested residues of the blank group(g)
Wre Weight of digested residues of the experimental group (g)
λ Lag phase time (d)
β0 Final biogas production (mL/gSV)
β (t) Cumulative biogas yield (mL/gSV)
µm Maximum biogas production rate (mL/gSV.d)

anaerobic microorganisms for biogas production (Chen et al., 2015). However, the efficient anaerobic digestion process impli-
cates the variables monitoring the operational conditions (temperature, pH, inoculum source, organic load, carbon/nitrogen
ratio) and the use of physical, chemical and/or biological pretreatments in to enhance the degradation of lignocellulosic
structure within rice straw. The latter is due to hemicellulose and lignin can be resistant to enzymatic action of anaerobic
consortium (Zheng et al., 2014). Given the relevance in the intrinsic characteristics of rice straw, including a recalcitrant
lignocellulosic structure and a high carbon/nitrogen ratio (C/N), several studies focus on pretreatment (Chen et al., 2014) and
an improvement of the degradation process have been conducted (Zhang et al., 2016; Gu et al., 2015; Li et al., 2015; Lei et al.,
2010). For instance, the hydrothermal treatment with water at 200 ◦ C has shown to significantly improve the degradation
process, allowing yields of methane up to 315.9 L/kgVS (Chandra et al., 2012), while physical extrusion produced yields up
to 227.3 L/kgVS (Chen et al., 2014). This last physical pretreatment can modify the straw properties as porosity, bulk density
and specific surface area. Chemical pretreatments with Ca(OH)2 allow to improve the hydrolysis of rice straw, increasing
the conversion of glucose and xylose (a predominant component of hemicellulose) and the biogas rate production by 574.5
L/kgVS (Gu et al., 2015). Furthermore, biological pretreatments with white mushrooms have shown an enzymatic action
on the lignocellulosic material to improve its subsequent anaerobic degradation, and increasing the biogas productions
(Rouches et al., 2016).
To increase the biogas yield, co-digestion of rice straw with animal manure is considered a more effective method for
C/N regulation, compared to the addition of chemical reagents. Li et al. (2015) investigated the effects of feedstock ratio
and organic loading rate (OLR) on the anaerobic mesophilic co-digestion of rice straw and cow manure in batch tests and
continuous bench experiments. They found that the optimal volatile solid ratio to be 1:1 (rice straw: cow manure) with
 C. Candia-García et al. / Environmental Technology & Innovation 10 (2018) 215–222 217

Table 1
Solid loading for first and second experimental runs.
First Experimental Run (FER) Reactors 101–102 Reactors 201–202 Reactors 301–302 Reactors 401–402
I/S Ratio (based on VS) 0.8 0.5 0.3 ∼
Rice Straw Mass (g) 6.5 10.4 17.3 ∼
Solid Loading Rate (gST/L mixture) 21.7 34.7 57.8 17.6
Second Experimental Run (SER) Batch reactors inoculated with the optimum I/S ratio
I/S Ratio 0.8
Rice Straw Mass (g) 6.5

a stable and efficient co-digestion process. However, various microbial sources from animal manure have been tested for
anaerobic degradation of lignocellulosic material (Gu et al., 2014; Wang et al., 2012). An appropriate selection of the microbial
communities can effectively improve the biomass biodegradability and methane yield. Several studies (Creevey et al., 2014;
Yue et al., 2013) showed that, rumen fluid, formed in the fore stomach of cows that included a complex microbial population.
It showed a higher ability and activity to degrade lignocellulosic biomass than other anaerobic microorganisms. In addition,
O’Sullivan et al. (2006) reported that the cellulose solubilization, by rumen microorganisms, is significantly faster than that
by microbial communities from landfills or anaerobic digesters. Despite this fact, there is a likelihood of 10 to 20% of the
rumen microbial population to be identified (Yue et al., 2013). Overall, a total of 20 bacterial phyla have been detected in the
rumen, but the majority of these are rare and only nine phyla have cultured representatives (Creevey et al., 2014).
This paper presents a comprehensive overview of anaerobic co-digestion on rice straw and rumen fluid under room
temperature. It evaluates the biogas production and determines the lignocellulosic composition changes, along retention
time with the aim to find an environmentally-friendly alternative for straw disposition in rural areas.

2. Methods

2.1. Feedstock and inoculum

Four representative varieties of rice straw were collected from rice fields in Saldaña, a town located in the state of Tolima–
Colombia. Varieties harvested were: Fedearroz 67 (F67), Fedearroz 733 (F733), Fedearroz 2000 (F200) and Fedearroz 768
(F768). These varieties were sun-dried and stored in black plastic bags at room temperature for about 15 days. After, they
were chopped with a food processor in sections 5 to 15 mm, following the methodology used by Chen et al. (2015).
Rumen content collected from the Slaughter Plant (Carlima), located in the city of Ibagué-Tolima, was used as the
inoculum source. All slaughtered cattle are quarantined and are not fed for 3 days before being processed. The inoculum
was stored in a plastic bottle of 5 L for about one month at room temperature (25 ◦ C to 27 ◦ C), and in absence of O2 .
Physicochemical composition analyses showed that the rumen content had a high humidity level (93.58%) and 6.42% of
total solids on wet basis. The rumen fluid presented a high concentration of total nitrogen (TN, 215 ± 23 mgN/L) and NH4 -N
(98 ± 15 mgN/L), thus it could serve as nitrogen source during methanogenic process without the addition of extra nitrogen
from other sources. The microbial community of the rumen fluid was not identified in this study.

2.2. Experimental set-up

Two experimental assays were carried out in this study. The assays were performed in batch reactors of 500 mL, with a
load volume of 300 mL, at room temperature (25 ◦ C to 27 ◦ C). A blank (rumen content alone) was carried out for each run
to correct endogenous methane production. Biogas production was measured daily and for 60 days of retention time, based
on other research models (Undersander and Mertens, 1993). In all cases, the batch reactors were run in duplicate.
The first experimental assay (FER) was conducted to determine the best Inoculum/Substrate ratio (I/S). Three different I/S
ratios (0.8, 0.5 and 0.3), based on volatile solids (VS) were tested. Next, the biogas production was measured daily for 60 days.
In the second assays (SER), 14 batch reactors were inoculated with the best I/S ratio obtained in FER assay. Henceforward,
cellulose, hemicellulose, lignin, cellular content and volatile solid (VS) concentrations were analyzed for 60 days. Reactors,
in the second experimental, run were purged with nitrogen and preheated at 45 ◦ C for 20 min to activate bacterial action.
Table 1 shows the solid loading rate for both experimental runs.

2.3. Analytical methods

Composite samples were analyzed for various physicochemical parameters, as Total Solids (TS), Volatile Solids (VS),
moisture, ash and Chemical Oxygen Demand (COD), according to Standard Methods for the Examination of Water and
Wastewater (APHA, 1998). Additionally, concentration of cellulose, hemicellulose, lignin, cellular content and silica, fol-
lowing regulatory methods for Detergent Fiber Neutral (DFN), Acid Detergent Fiber (ADF) and Detergent Lignin Fiber (LDF)
specified by Undersander and Mertens (1993).
Biogas production was performed using water displacement equipment, with a measure range of 1 to 100 mL of
biogas. This equipment was built taking into account the principles of fluid pressure and the model suggested by Ojikutu
218 C. Candia-García et al. / Environmental Technology & Innovation 10 (2018) 215–222

and Osokoya (2014). Biogas composition was determined according to headspace static technique and analysis by gas
chromatography with thermal conductivity detector and flame ionization detector (S-HS/GC/TCD/FID). The chromatographic
analysis was performed with a gas chromatograph AT 6890A from Agilent Technologies Company. Gas analysis was made
by static headspace HP7694E machine from Hewlett-Packard Company.

2.3.1. Degradation efficiency

The degradation efficiency of the different substance composing the rice straw was monitored for 60 days of retention
time. Removal calculations for total solid (TS), volatile solid (VS), cellulose, hemicellulose, lignin, silica and cellular content
(Eqs. (1) to (6)), were made according to Chen et al. (2014). The list of variables in each equation is shown in initial
nomenclature table.
SLR × VW × TSrs − Wre × TSre − Wb × TSb
VSde = (1)
SLR × VW × TSrs
SLR × VW × TSrs × VSrs − Wre × TSre × VSre − Wb × TSb × VSb
VSde = (2)
SLR × VW × TSrs × VSrs
SLR × VW × TSrs × Cellulosers − Wre × TSre × Cellulosere
Cellulosede = (3)
SLR × VW × TSrs × Cellulosers
SLR × VW × TSrs × Hcellulosers − Wre × TSre × Hcellulosere
Hcellulosede = (4)
SLR × VW × TSrs × Hcellulosers
SLR × VW × TSrs × Ligninrs − Wre × TSre × Ligninre
Ligninde = (5)
SLR × VW × TSrs × Ligninrs
SLR × VW × TSrs × CellularC .rs − Wre × TSre × CellularC .re
CellularC .de = . (6)
SLR × VW × TSrs × CellularC .rs

2.3.2. Kinetic modeling

The modified Gompertz model was applied to adjust the experimental biogas production Eq. (7). This model has been
applied in modeling methane production in the anaerobic digestion process (Li et al., 2013).
µm e
{ [ ]}
β (t) = β0 exp − exp (λ − 1) + 1 (7)
β0
where β (t) is the cumulative biogas yield (mL/gSV), β0 is the final biogas production (mL/gSV), µm refers to the maximum
biogas production rate (mL/gSV.d), λ is the lag phase time (d), e is equal to 2.72 and t means the anaerobic digestion time (d).
Furthermore, the parameters (µm and λ) were estimated using a non-linear square method of Matlab (lsqnonlin function)
for all I/S ratios tested.

2.3.3. Statistical analysis

All the experiments were performed in duplicate, and the results were expressed as mean values and standard deviation.
Standard statistical procedures were used, including standard deviation, mean averaging and absolute difference. The
significance of variance test was determined using variance analysis with a significant level of 0.05 (p ≤ 0.05 considered
significantly different).

3. Results and discussion

3.1. Characterization of rice straw varieties

The chemical compositions of four varieties of rice straw used in Saldaña (Tolima) are shown in Table 2. Results obtained
were analyzed statistically and they show that there are not significant differences between analyzed rice straw varieties
(p > 0.05). TS, VS, ashes, cellulose, hemicellulose, lignin, cellular content and silica for the four varieties studied showed
similar behavior, with high values of volatile solids (82.51% to 84.37%), cellulose content (37.27% to 42.61%), and relevant
quantities of hemicellulose (10.12% to 12.55%) and lignin (9.26% to 11.89%). These results can classify the rice straw into
herbaceous biomass, that generally present cellulose contents higher than 32% and lignin contents lower than 19% (Xu et al.,
2014). Other researchers have reported similar high concentrations of volatile solids (70.37% to 83.18%), cellulose (25.4% to
35.5%), hemicellulose (16.70%) and lignin (10.4% to 11.2%) which represents an important fraction of rice straw composition
(Gu et al., 2014; Lei et al., 2010).
Overall, chemical composition and silicification are key factors that affect the utilization efficiency and nutritive quality
of lignocellulosic materials, as rice straw. According to Xu et al. (2014) the characteristics of lignocellulosic material are
correlated with methane yield; it is positive with cellulose content and very negative with extractives and lignin content.
Besides these feedstock characteristics, methane yield also relies on the actual operating conditions on anaerobic digestion.
For example, key factors that have been found to affect the performance of the anaerobic process include temperature, TS
content, carbon/nitrogen (C/N) ratio, particle size of the feedstock, concentration of inhibitors (e.g. ammonia), and alkalinity
of the system (Zhang et al., 2016; Gu et al., 2015; Li et al., 2015; Lei et al., 2010).
 C. Candia-García et al. / Environmental Technology & Innovation 10 (2018) 215–222 219

Table 2
Results of characterization of 4 rice straw varieties cultured in Saldaña–Tolima. Values in relation with dry matter.
Parameters Fedearroz 767 (F767) Fedearroz 733 (F733) Fedearroz 2000 (F2000) Fedearroz 768 (F768)
Volatile Solids (% TS) 82.51 ± 0.30 84.37 ± 0.36 83.45 ± 0.16 82.60 ± 0.24
Ashes (% TS) 17.49 ± 0.30 15.63 ± 0.36 16.55 ± 0.16 17.40 ± 0.24
Hemicellulose (% TS) 12.55 ± 2.33 15.35 ± 0.63 13.14 ± 1.04 10.12 ± 0.24
Cellulose (% TS) 40,12 ± 0,88 38.95 ± 0.77 37.27 ± 0.36 42.61 ± 0.24
Lignin (% TS) 11.89 ± 2.89 10.56 ± 0.31 11.27 ± 0.61 9.26 ± 0.24
Silica (% TS) 5.88 ± 0.29 5.92 ± 1.19 8.20 ± 1.11 6.67 ± 0.24
Cellular Content (% TS) 29.56 ± 0.61 29.22 ± 1.36 30.12 ± 1.18 31.34 ± 0.24

3.2. Biogas production

Three different I/S ratios (0.3; 0.5; 0.8) were monitored daily to determine the biogas production. The biogas curves are
shown in Fig. 1. Biogas production started after inoculation and continued to increase until they reached a peak (3 days)
for all assays, indicating proper initial condition for anaerobic digestion. Indeed, the assay with I/S ratio = 0.3 reached the
highest peak of daily biogas production with 146 ± 1 mL biogas in the second day. Although the biogas production for all
tests showed a high biogas generation in the first days, it can be seen that for ratios 0.5 and 0.3 biogas fell under 30 mL per
day, along the retention time, while reactors inoculated with I/S ratio = 0.8 showed steadier production for the 60 days.
Highest biogas production was obtained in I/S = 0.8 treatment with a biogas yield of 306 L/kgVS (FER) and 410 L/kgVS
(SER), with a methane content over 70% (Fig. 2). Other researchers reported similar results of biogas yield. Gu et al. (2014)
reported a biogas production of 325.3 L/kgVS using rice straw and dairy manure as inoculum source with an I/S ratio of
0.5. Lei et al. (2010) described a maximum biogas production of 350 L/kgSV with a chemical pretreatment using phosphate
supplement. Amnuaycheewa et al. (2016) showed that pretreatments with organic acids can provide biogas yields from
322.1 L/kgVS.
Nevertheless, reactors inoculated with I/S = 0.3 and 0.5 ratios, produced 30 and 118 L/kgVS, respectively. Best treatment
(I/S = 0.8) exhibits a continuous and marked growth slope along the retention time in comparison with I/S = 0.3 and 0.5
treatments. It showed an apparent stabilization of slope after 20 days of anaerobic digestion. Results showed that cumulative
biogas yield was influenced by I/S ratio. Indeed, the increase of microorganisms (inoculum) favors the biodegradation of the
lignocellulosic material. These results coincide with Xu et al. (2014), in anaerobic degradation of lignocellulosic material, the
decrease of I/S ratio significantly reduces methane yield, which is probably due to a reduction in microbial population and
alkalinity at low I/S ratio. Furthermore, an increase of substrate could cause an overloading or substrate inhibition process
by the accumulation of volatile fatty acids (VFAs) which indicate an inadequate balance between hydrolysis, acidogenesis
and methanogenesis stages. According to our assays, the best results can be obtained at I/S ratios close to 1, which can be
analogous to co-digestion methods (Li et al., 2015). Co-digestion is related as a great alternative to treat multiple wastes
getting more balanced nutrients and ideal Carbon/Nitrogen ratios (C/N), which provides synergistic and complementary
effects that allow best methane yields (Chen et al., 2015). This aspect was evidenced in this study, as shown in Fig. 2, where
the cumulative biogas yield increased with increase the I/S ratios.
Other studies have shown that particle size is in an inverse correlation with methane yield (Xu et al., 2014). This
is acceptable in view of small particle size providing high specific surface area favorable for digestion anaerobia of
lignocellulosic biomass. Although, in some literature small particle size can cause excessive hydrolysis, leading to quick
volatile fatty acids (VFA) accumulation (Zheng et al., 2014). However, the reduction of particle size in this study (5 to 15 mm)
did not cause inverse effects.
As suggested in the above, co-digestion could be an interesting technology for treatment of rice straw and rumen.
Anaerobic co-digestion is increasingly used to simultaneously treat organic wastes, in order to better nutrient balance,
improve digester operational advantages and co-substrate handling, reduce the negative effects of toxic compounds on
the process, and increase the biogas yield. Moreover, co-digestion technology contributes to used multiple streams of
waste can be processed in a single plant of anaerobic digestion at the same time. However, for a full-scale biogas plant
is necessary to guarantee the availability of suitable amounts of the different kinds of substrate for co-digestion, because a
change to the input parameters, as a type of feedstock in the biogas system, may affect the operating costs and total energy
production. In addition, the application of co-digestion could cause additional efforts due to additional mixing requirements,
homogenization, heat treatment, biodegradation time, etc. Nonetheless, future development a biogas plant of co-substrates
digestion (rice straw and rumen) requires a more detailed study, long-term evaluation in pilot plant and a careful research
for a sustainable operation.

3.3. Kinetic modeling

The Gompertz model is useful to explain lag time and sigmoidal growth curve of biogas production from anaerobic
degradation of lignocellulosic material (Fig. 2). The squared correlation coefficient, R2 , was close to unity for I/S ratio cases =
0.8, indicates a good fit in describing the biogas production. However, in other two cases (I/S = 0.3 and 0.5), the R2 coefficients
were under 0.6, which revealed that the biogas production is over predicted in the model for both cases. These results suggest
220 C. Candia-García et al. / Environmental Technology & Innovation 10 (2018) 215–222

Fig. 1. Daily biogas production for treatments in first experimental run (FER) and second experimental run (SER).

Fig. 2. Cumulative biogas yield for different I/S ratios. The Gompertz model is the solid line.

that for an optimal anaerobic degradation of rice straw with rumen as inoculum, I/S ratio could be close to 1. This may be
due to high C/N ratio of rice straw; it could cause a lack of nitrogen and nutrient. Indeed, Zhang et al. (2016) showed a C/N
ratio of 64.1 for rice straw, thus, rice straw is not ideal, as the sole feedstock for anaerobic digestion. This result coincides
with previous studies that reported a low correlation between C/N ratio and methane yield when the value of the C/N (range
from 18 to 30). The latter is regarded as suitable for anaerobic digestion of lignocellulosic material (Xu et al., 2014). In such
condition, the C/N ratio might not strongly affect methane yield. Likely, the most favorable C/N ratio in our assays was
achieved with I/S ratio = 0.8, while for the other assays C/N ratio (I/S = 0.3 and 0.5) showed a strong influence on biogas
production.
According to the results, the lag phase time estimate (λ) was 3.01 and 4.68 days and the maximum biogas production rate
(µm ) was 7.49 and 9.56 mL/gSV.d, for I/S ratios = 0.8 in the two tests, FER and SER assays, respectively. Similar results were
obtained by Zhang et al. (2016). They showed 10.66 mL/gSV.d and 3.21 of µm and λ, respectively; for methane production
of rice straw with rumen (I/S = 1, without pretreatment) under mesophilic condition. Therefore, the parameter values
dependent of I/S ratio and an optimal degradation relied on the C/N ratio. Indeed, a lower λ and a higher µm can mean a faster
startup and a higher efficiency of anaerobic digestion process (Zhang et al., 2016). These modeling results also indicate that
the co-digestion of rice straw and rumen fluid could significantly enhance biogas production rate when the lag phase time
is shortened and the methane yield is increased. Therefore, this condition might be obtained with a suitable pretreatment.

3.4. Degradation efficiency and organic matter removal

Degradation efficiency and organic matter removal were determined in a reactor with I/S ratio of 0.8 (SER). Fig. 3 exhibits
the results obtained during 60 days of rice straw biodegradation under anaerobic condition and pH of about 6.5. The removals
 C. Candia-García et al. / Environmental Technology & Innovation 10 (2018) 215–222 221

Fig. 3. Degradation efficiency of organic material from rice straw along retention time.

Fig. 4. Concentration changes in compositional structures of rice straw along retention time.

were determined in terms of TS (32.62%), VS (41.23%), cellulose (50.10%), hemicellulose (44.17%) and lignin (32.62%), in
relation to initial composition of rice straw during a 60 day-treatment. According the Fig. 3, the maximum cellulose removal
was obtained after 40 days of treatment; similar results were observed for TS, VS and lignin. However, the hemicellulose
removal continues with a slight increase over 60 days. Results suggest that the lignin structure showed a lower degradation,
in comparison with cellulose and hemicellulose structure, whose biodegradation mechanisms have been studied in other
researches and predicted higher yields of biofuel by destructing resistant enzymatic structures as lignin Arantes et al. (2011).
However, notable lignin degradation in this study was found in the co-digestion process, suggesting the possible presence
of lignin degrader in the rumen fluid. Our results were higher than those shown by Zhang et al. (2016), who obtained 20.6%
of lignin degradation efficiency after a biological pretreatment using rumen fluid. Indeed, the rumen microorganisms can
degrade the lignin of rice straw because the hydrolyzing enzymes could pass through the holes on the fiber surface (Yue
et al., 2013). Moreover, cellulose, hemicellulose and lignin are the main carbon source for anaerobic community and the
availability of a simpler source of carbon (cellulose and hemicellulose) would affect the subsequent lignin degradation.
To our knowledge, the lignocellulosic biomass is a heterogeneous complex of carbohydrate and polymers. Cellulose is
the most abundant polysaccharide in herbaceous biomass (rice straw, corn stover, wheat straw, and switch grass) and hemi-
cellulose is a heteropolysaccharide with different monosaccharides, as pentoses and hexoses where xylose is predominant
(Himmel et al., 2007). The ramified structure makes hemicellulose more resistant than cellulose to hydrolyzation (Mosier
et al., 2005), while lignin is a polymer containing phenol strongly linked to polysaccharides. They enhance the resistance of
fibers to microbial attack (Himmel et al., 2007). In fact, previous studies reported that the lignin determines the anaerobic
biodegradability degree of substrates and a high lignin content is a negative indicator for methane production (Xu et al.,
2014; Buffiere et al., 2006).
Fig. 4 shows the concentration behaviors for organic and inorganic changes in lignocellulosic matter during anaerobic
digestion treatment. It is possible to see that the fraction of VS is composed by cellulose, hemicellulose, lignin and a fraction
of cellular content, while an inorganic fraction is represented by a little part of cellular content and silica. Silica has been
described as a characteristic structure from rice plant which allows to keep energy by replacing lignin for silica, as part of
an evolutionary process adapted from many plants (Van Soest, 2006). The final removal of total matter at day 60 was 10.47
gST/L (40%). These changes represent the conversion process in anaerobic digestion where part of volatile solids in loading
rate is converted to biogas. In fact, biogas is generated from biological conversion of substrate. In our case, substrate can be
represented by TS and VS, which can become more available by anaerobic microorganisms throughout the process.

4. Conclusion

The results obtained in this study clearly demonstrate that the anaerobic digestion is a promising alternative to treat the
sub-products of rice crop, as rice straw. The efficient biodegradation of rice straw via anaerobic digestion relies on of I/S
222 C. Candia-García et al. / Environmental Technology & Innovation 10 (2018) 215–222

ratios. Indeed, the best biogas yield was obtained at I/S ratio close to 1, which can be analogous to co-digestion methods.
In this method, it is possible to get a more balance nutrient and an ideal C/N ratio for anaerobic digestion because the high
C/N ratio of rice straw limits the anaerobic degradation, as waste is only used as feedstock of the process. The experimental
data also demonstrate the efficient removal of organic matter and lignocellulosic material reaching values of 50.10% for
cellulose, 44.17% for hemicellulose and 32.62% for lignin. Furthermore, it was demonstrated that the use of natural microbial
consortium, as rumen fluid for lignocellulosic material degradation could be effective and a promising option.

Acknowledgments

This project, identified with the code number 15-334-SEM, was funded by the Universidad de Ibagué and the Gobernación
del Tolima through the agreement number 1026. The authors wish to thank Carlima for providing the rumen content.

References

Agencia de Noticias UN, 2014. Con hongos degradan y reutilizan desechos del arroz, WWW Document. Univ. Nac. Colomb. URL http://agenciadenoticias.
unal.edu.co/detalle/article/con-hongos-degradan-y-reutilizan-desechos-del-arroz.html (Accessed 13 June 2016).
Amnuaycheewa, P., Hengaroonprasan, R., Rattanaporn, K., Kirdponpattara, S., Cheenkachorn, K., Sriariyanun, M., 2016. Enhancing enzymatic hydrolysis and
biogas production from rice straw by pretreatment with organic acids. Ind. Crops Prod. 87, 247–254. http://dx.doi.org/10.1016/j.indcrop.2016.04.069.
APHA, 1998. Standard methods for the examination of water and wastewater.
Arantes, V., Milagres, A.M.F., Filley, T.R., Goodell, B., 2011. Lignocellulosic polysaccharides and lignin degradation by wood decay fungi: the relevance of
nonenzymatic Fenton-based reactions. J. Ind. Microbiol. Biotechnol. 38, 541–555. http://dx.doi.org/10.1007/s10295-010-0798-2.
Buffiere, P., Loisel, D., Bernet, N., Delgenes, J.P., 2006. Towards new indicators for the prediction of solid waste anaerobic digestion properties. Water Sci.
Technol. 53, 233–241.
Cardona, L.E., 2015. Estos son los desafíos de los productores de arroz en el país. El País.
Chandra, R., Takeuchi, H., Hasegawa, T., 2012. Hydrothermal pretreatment of rice straw biomass: A potential and promising method for enhanced methane
production. Appl. Energy 94, 129–140. http://dx.doi.org/10.1016/j.apenergy.2012.01.027.
Chen, X., Yuan, H., Zou, D., Liu, Y., Zhu, B., Chufo, A., Jaffar, M., Li, X., 2015. Improving biomethane yield by controlling fermentation type of acidogenic phase
in two-phase anaerobic co-digestion of food waste and rice straw. Chem. Eng. J. 273, 254–260. http://dx.doi.org/10.1016/j.cej.2015.03.067.
Chen, X., Zhang, Y., Gu, Y., Liu, Z., Shen, Z., Chu, H., Zhou, X., 2014. Enhancing methane production from rice straw by extrusion pretreatment. Appl. Energy
122, 34–41. http://dx.doi.org/10.1016/j.apenergy.2014.01.076.
Creevey, C.J., Kelly, W.J., Henderson, G., Leahy, S., 2014. Determining the culturability of the rumen bacterial microbiome. Microb. Biotechnol. 10, 1751–7915.
Gu, Y., Chen, X., Liu, Z., Zhou, X., Zhang, Y., 2014. Effect of inoculum sources on the anaerobic digestion of rice straw. Bioresour. Technol. 158, 149–155.
http://dx.doi.org/10.1016/j.biortech.2014.02.011.
Gu, Y., Zhang, Y., Zhou, X., 2015. Effect of Ca(OH)2 pretreatment on extruded rice straw anaerobic digestion. Bioresour. Technol. 196, 116–122. http:
//dx.doi.org/10.1016/j.biortech.2015.07.004.
Himmel, M.E., Ding, S.Y., Johnson, D.K., Adney, W.S., Nimlos, M.R., Brady, J.W., Foust, T.D., 2007. Biomass recalcitrance: engineering plants and enzymes for
biofuels production. Science 315, 804–807.
Lei, Z., Chen, J., Zhang, Z., Sugiura, N., 2010. Methane production from rice straw with acclimated anaerobic sludge: Effect of phosphate supplementation.
Bioresour. Technol. 101, 4343–4348. http://dx.doi.org/10.1016/j.biortech.2010.01.083.
Li, Y., Feng, L., Zhang, R., He, Y., Liu, X., Xiao, X., Ma, X., Chen, C., Liu, G., 2013. Influence of inoculum source and pre-incubation on bio-methane potential of
chicken manure and corn stover. Appl. Biochem. Biotechnol. 171, 117–127. http://dx.doi.org/10.1007/s12010-013-0335-7.
Li, D., Liu, S., Mi, L., Li, Z., Yuan, Y., Yan, Z., Liu, X., 2015. Effects of feedstock ratio and organic loading rate on the anaerobic mesophilic co-digestion of rice
straw and cow manure. Bioresour. Technol. 189, 319–326. http://dx.doi.org/10.1016/j.biortech.2015.04.033.
Mosier, N., Wyman, C., Dale, B., Elander, R., Lee, Y.Y., Holtzapple, M., Ladisch, M., 2005. Features of promising technologies for pretreatment of lignocellulosic
biomass. Bioresour. Technol. 96, 673–686.
Ojikutu, A., Osokoya, O., 2014. Evaluation of biogas production from food waste. Internat. J. Engrg. Sci. 3, 01–07.
O’Sullivan, C.A., Burrell, P.C., Clarke, W.P., Blackall, L.L., 2006. Comparison of cellulose solubilization rates in rumen and landfill leachate inoculated reactors.
Bioresour. Technol. 97 (18), 2356–2363.
Rouches, E., Herpoël-Gimbert, I., Steyer, J.P., Carrere, H., 2016. Improvement of anaerobic degradation by white-rot fungi pretreatment of lignocellulosic
biomass: A review. Renew. Sustain. Energy Rev. 59, 179–198. http://dx.doi.org/10.1016/j.rser.2015.12.317.
Undersander, D., Mertens, D.R., 1993. Forage analyses procedures.
Van Soest, P.J., 2006. Rice straw, the role of silica and treatments to improve quality. Anim. Feed Sci. Technol. 130, 137–171. http://dx.doi.org/10.1016/j.
anifeedsci.2006.01.023.
Wang, X.J., Yang, G.H., Feng, Y.Z., Ren, G.X., Han, X.H., 2012. Optimizing feeding composition and carbon-nitrogen ratios for improved methane yield during
anaerobic co-digestion of dairy, chicken manure and wheat straw. Bioresour. Technol. 120, 78–83.
Xu, F., Wang, Z.-W., Li, Y., 2014. Predicting the methane yield of lignocellulosic biomass in mesophilic solid-state anaerobic digestion based on feedstock
characteristics and process parameters. Bioresour. Technol. 173, 168–176. http://dx.doi.org/10.1016/j.biortech.2014.09.090.
Yue, Z.B., Li, W.W., Yu, H.Q., 2013. Application of rumen microorganisms for anaerobic bioconversion of lignocellulosic biomass. Bioresour. Technol. 128,
738–744.
Zhang, H., Zhang, P., Ye, J., Wu, Y., Fang, W., Gou, X., Zeng, G., 2016. Improvement of methane production from rice straw with rumen fluid pretreatment: A
feasibility study. Int. Biodeterioration Biodegrad. 113, 9–16. http://dx.doi.org/10.1016/j.ibiod.2016.03.022.
Zheng, Y., Zhao, J., Xu, F., Li, Y., 2014. Pretreatment of lignocellulosic biomass for enhanced biogas production. Prog. Energy Combust. Sci. 42, 35–53.
http://dx.doi.org/10.1016/j.pecs.2014.01.001.