Journal of Cleaner Production 213 (2019) 176e184

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Journal of Cleaner Production

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

Production of biogas and biofertilizer using anaerobic reactors with

swine manure and glycerin doses
Maria Luiza Veroneze a, Daniel Schwantes b, *, Affonso Celso Gonçalves Jr. c, **,

ssica Manfrin b, Andre
Alfredo Richart d, Je
ia da Paz Schiller b, Thiago Bana Schuba a
a
State University of Western Parana
, Postgraduate Program in Energy Engineering in Agriculture, Campus of Cascavel, Universita

ria Street, 2069, Jardim

rio, Cascavel, Parana
Universita
State, 85819-110, Brazil
b
Educational College of Medianeira, 1820 Rio Branco Street, Downtown, Medianeira, State of Parana
, 85884-000, Brazil
c
State University of Western Parana
, Postgraduate Program in Agronomy, Campus of Marechal Ca ^ndido Rondon, Pernambuco Street, 1777, Centro, Marechal
Ca^ndido Rondon, Parana
State, 85960-000, Brazil
d
Pontifical Catholic University of Parana
, Agronomy, Campus of Toledo, Union Avenue, 400, Jardim Coopagro, Toledo, Parana State, 85902-532, Brazil

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

Article history: This research aims to evaluate the development of biodigestion of swine manure with addition of
Received 25 March 2018 glycerin doses, in order to promote a cleaner destination for both of these wastes in agroindustry,
Received in revised form adopting different concentrations for a better relation of biogas production and biostabilized fertilizer.
14 December 2018
For this, five anaerobic biodigestors were developed, each reactor was fed with swine manure consid-
Accepted 18 December 2018
Available online 18 December 2018
ering four treatments (5%, 10%, 15% and 20% vv
1 of glycerin). The physical-chemical parameters (tem-
perature, pH, turbidity, solids series, total phosphorus, total nitrogen (Kjeldahl), and COD were evaluated
every seven days, moreover the biogas production was monitored daily by a gasometer. The concen-
Keywords:
Renewable energy
trations of Cu, Zn, Mn, Fe, Cd, Pb and Cr during hydraulic retention time (HRT) were determined by Flame
Biofuel Atomic Absorption Spectrometry (FAAS). Anaerobic co-digestion of swine and glycerin wastes didn't
Glycerin provided higher biogas production than the biodigestion without the addition of glycerin. The biogas
Co-digestion production rates were impaired by the addition of glycerin doses higher than 5%, and the results
Biogas production demonstrate that the nutrient concentrations in the biostabilized waste are expressive for vegetal bio-
fertilization, mainly for N, P, K, Ca, Mg and micronutrients such as Cu and Zn.
© 2018 Elsevier Ltd. All rights reserved.

1. Introduction treating them as waste production (Kolesa
rova
et al., 2011).

Nowadays, one of the most important issues in different sectors
is to develop new technologies and process that ensure the sus-
tainable development (Dovì et al., 2009). In this way, renewable
energy sources and biofuels, including biodiesel, are receiving
increasing attention as substitutes for fossil fuels. However, its
implementation in the market depends mainly on making these
sources more competitive. An option to reduce the costs of biofuels
is to use co-products as a potential source of energy instead of
* Corresponding author.
** Corresponding author.
E-mail addresses: marial_veroneze@hotmail.com (M.L. Veroneze), daniel_
schwantes@hotmail.com (D. Schwantes), affonso133@hotmail.com
(A.C. Gonçalves), alfredo.richart@pucpr.br (A. Richart), jessicamanfrinn@gmail.
com (J. Manfrin), andreia.schiller@hotmail.com (A. da Paz Schiller), t.schuba@
hotmail.com (T.B. Schuba).
Launched on December 6, 2004 by the Federal Government of
Brazil, the “Brazilian Program for the Production and Use of Bio-
diesel” is based on the growing demand for renewable fuels and the
Brazilian potential to meet a significant portion of these needs.
However, the introduction of biodiesel in the Brazilian energy
matrix raises the quantities of its main co-product, which is glyc-
erin, a co-product generated at the rate of 10 kg for every 100 L of
biodiesel (Larsen et al., 2013).
In 2016, according to the National Petroleum Agency, Brazil
produced 430,807 kg of glycerin (equivalent to 341.9 m3) (ANP,
2016), although glycerin has a variety of uses in multiple indus-
trial sectors, the increasing production of biodiesel may cause
excess glycerin, reducing its price, and possible causing environ-
mental problems.
Considering that glycerin is transformed into several products,
such as pharmaceuticals and cosmetics, however the enormous
amount of this co-product can exceed the absorption capacity of

https://doi.org/10.1016/j.jclepro.2018.12.181
0959-6526/© 2018 Elsevier Ltd. All rights reserved.
 M.L. Veroneze et al. / Journal of Cleaner Production 213 (2019) 176e184
the national market (Leonel et al., 2012), and that the production of
biodiesel production from soybean in Brazil is responsible to many
environmental impacts (Cavalett and Ortega, 2010) justifying
studies that apply this substance for other purposes as a new
alternative for the energy sector, improving this department.
Some research has already shown that anaerobic digestion of
crude glycerin (GC), with different wastes and effluents, is an
excellent option to increase the production of biogas and methane,
improving the treatment of these materials (Orrico Jr. et al., 2016).
In this context, recent research has tested the capacity of using
bovine and swine manure associated with crude glycerin, aiming
the stabilization of organic matter and the production of biogas
(Schwingel et al., 2016; Simm et al., 2017).
These studies become of fundamental importance, since Brazil is
a growing economy, based on agribusiness (Schwingel et al., 2016),
and is the fourth largest producer and exporter of meat from swine in
the world, which is responsible for the generation of large waste
amounts (Cherubini et al., 2015). . Swine production generates large
amounts of manure, and this is the main environmental problem of
this activity, mainly due to the high concentrations of organic matter,
nutrients and metals of its constitution (Orrico Júnior et al., 2010).
The swine wastes are mainly used for agriculture purposes, in
biofertilizer form, however, their inadequate management can lead
to the contamination of water courses, groundwater, soil and air,
representing a risk to the sustainability and expansion of swine
farming as an economic activity (Kunz et al., 2015).
Among the methods of waste treatments most used in pig
farming, it is highlighted the use of pond systems, biodigestion
processes and others. These alternatives promote economic and
ecological sustainability.
In this context, the process of biodigestion of swine manure and
glycerin, which have different origins and compositions, can be
characterized as a possible solution to improve the treatment and
biostabilization of these residues, possibly increasing biogas
production.
The objective of the present study was to evaluate the digestion
process of swine manure and doses of glycerin by using alternative
biodigesters, through the determination of the nutrient contents in
the biostabilized effluent and biogas productivity.
2. Material and methods
2.1. Development of alternative anaerobic reactors
The biodigestors were constructed with recycled plastic mate-
rial, for that, five plastic bottles with capacity of 50 L were used; 5
A B
Fig. 1. Biodigesters and gasometer linked to the used reactors.
177
taps; 4.5 m of hose Rodoar soft blue PU 8 mm; 5 metal and silicone
fittings used for sealing, keeping the internal environment
completely anaerobic.
The taps were installed to allow sampling of the liquid effluent
along the hydraulic retention time (HRT), the reactors were ar-
ranged horizontally, with the end containing the tap down and a
spigot, facing upwards (Fig. 1A).
2.2. Installation of the experiment and glycerin doses
The experiment was carried out at the Pontifical Catholic Uni-
versity of Paran
a (PUCPR), located in the municipality of Toledo,
Parana
(24 430 08.7200 S and 53 460 08.7200 W) with an average
elevation of 575 m.
Before the supply of the biodigestors, physical and chemical
parameters of swine manure were analyzed, in order to evaluate
the efficiency of the process.
The exact volume of 25 L of effluent were added in each bio-
digester, as well as increasing doses of glycerin (5, 10, 15 and 20%
vv
1). The biodigesters were operated without maintenance of
temperature, with hydraulic retention time (HRT) of 42 days, with
daily monitoring of gas volume and weekly monitoring of physical
and chemical parameters on the effluent.
In Table 1 are exhibited the data regarding the amount of swine
manure and glycerin (Vetec®, 99.5%) added to the biodigesters.
2.3. Gasometer
Five gasometers were made in plastic bottles, which were filled
with water and sealed with a lid to which two hoses were con-
nected, one for biogas inlet and another for the water outlet. As the
gas was produced, it was directed through the hoses for the bottles,
the volume of biogas produced ended up generating pressure and
displacing the water to a second bottle, thus providing quantifica-
tion of the produced gas volume (Fig. 1B).
For the construction of the gasometers, 10 plastic bottles of 2 L
were used; 4.5 m of hose Rodoar soft blue PU 8 mm; 5 metal fit-
tings; 5 plastic valves and silicone for sealing.
2.4. Waste sampling and evaluated parameters
Sampling of swine manure was carried out in a property located
a (24.77066000 S and 53.69516900
in the municipality of Toledo, Paran

W), the property had 910 pigs, with 25 days of life being used for
termination.
Physical and evaluated chemical parameters were: pH;
178 M.L. Veroneze et al. / Journal of Cleaner Production 213 (2019) 176e184

Table 1 for the satisfactorily biodigestion process, especially if the pH drop

Doses of glycerin and total volume of biodigesters. occurs rapidly in the initial phases of anaerobic fermentation
Doses of glycerin Swine manure Glycerin Total volume (Schwingel et al., 2016).
% (vv
1) volume (L) volume (L) of mixture (L) The pH value of the substrate may influence the performance of
0 25.00 0.00 25.00 the steps when it is too much alkaline or acid, because although the
5 23.75 1.25 25.00 microorganisms act in different pH ranges depending on the bio-
10 22.50 2.50 25.00 digestion phase, the populations of methanogenic microorganisms
15 21.25 3.75 25.00
responsible for the production of biogas, are quite sensitive to
20 20.00 5.00 25.00
variations in pH.
According to Calli et al. (2005), in an biodigestion process it is
possible that inhibition of biogas production occurs at a pH higher
than 8.3. Ferrer et al. (2010) mention that the pH range between 6.1
turbidity; total (TS), fixed (FS) and volatile (VS) solids; total phos-
and 8.3 can be considered acceptable for biodigestion.
phorus (Ptotal); total nitrogen (NKjeldahl) (Ntotal), and chemical oxy-
This relationship was also observed by Holm-Nielsen et al.
gen demand (COD) (APHA, 2012).
(2008), when studying the fermentation of waste with organic
In the liquid phase, the total concentration of potassium (K),
compounds from food industry and pure glycerol (99.5% glycerol).
calcium (Mg), copper (Cu), zinc (Zn), iron (Fe), manganese (Mn),
In this case the inclusion of 3.5e6.5% of glycerol, in relation to the
cadmium (Cd), lead (Pb) and chromium (Cr) were determined by
total volume of the digester, promoted greater production of vol-
Flame Atomic Absorption Spectrometry (FAAS) model GBC 932 AA
atile fatty acids, causing inhibition of the methanogenic stage and
(Victoria, Australia) with deuterium lamp for background correc-
subsequent collapse of the system.
tion (Welz and Sperling, 1999).
According to Rodrigues (2005), the rapid degradation of the
In addition, the volume of the produced biogas was determined
substrate may have provided an ideal environment for the bacteria
daily, by gasometers linked to the biodigesters, the measurement
that degrade the fibers, because after the consumption of glycerol
was carried out with graduated cylinder.
(present in the crude glycerin), ammonia is generated inside the
biodigesters, leading to acidification of the medium, which could
3. Results and discussion increase the population of bacteria of the acidogenic phase, which
produce organic acids.
3.1. Physical and chemical parameters of the manure in A rapid increase in turbidity (Fig. 3) was observed at dose 0%,
biostabilization followed by reduction of this parameter at the end of the 42 days of
evaluation. In addition, with glycerin additions (treatments 5, 10, 15
Fig. 2 shows the results of the determination of the pH of the and 20%) there is an initial decrease in turbidity and a small in-
effluent during the HRT in the biodigesters, as well as the average crease in the values at the end of HRT.
values of air temperature during the evaluation period. The biological reactions that occur in the anaerobic reactors are
It is possible to observe that acidification of the liquid medium related to the rapid and exponential reproduction of microorgan-
occurred as the glycerin doses increased, a fact evidenced during isms (catabolic reactions). Compared to the experimental data, it is
the course of HRT. This acidification of the medium through the observed that the doses with glycerin exhibited low turbidity when
elevation of the glycerin doses results from the microbial meta- compared to the dose 0%, which can be indicative of the biological
bolism in the biodigesters, because in organic substrates of rapid activity in each evaluated reactor, since the larger the population of
degradation, there is also a rapid increase in the population of microorganisms in suspension the greater the turbidity of the
bacteria (Simm et al., 2017). medium.
This result suggests that in glycerin treatments there was a Another important fact concerns the environment of the
higher production of short chain fatty acids, which are fundamental

7.0 35.0

6.5
Air Temperature (oC)

30.0

6.0
25.0
5.5
pH

20.0
5.0

15.0
4.5

4.0 10.0
7 14 21 28 35 42

HRT (days)

Max air temp. Average air temp. Min. ar temperature Dose 0%

Dose 5% Dose 10% Dose 15% Dose 20%

Fig. 2. Average pH values for swine manure þ glycerin doses (% in vv
1), as well as average values for air temperature during the 42 days of evaluation. Note: pH of the swine
manure in HRT 0 days: 6.86 ± 0.12.
 M.L. Veroneze et al. / Journal of Cleaner Production 213 (2019) 176e184
Fig. 3. Average turbidity (NTU) in biodigesters by addition of glycerin doses (vv
1)
during 42 days of anaerobic biodigestion (a) and typical curve of bacterial growth (b)
(Persing et al., 2016). Note: Turbidity of swine manure in HRT of 0 days: 4170 UNT
±145.
microorganisms in the medium, since in dose 0% there is no change
in the medium, so that the adaptation phase of the microorganisms
in the medium has already occurred (Lag Phase shown in Fig. 3 b).
While in reactors with addition of glycerin, this adaptation appears
to occur throughout the HRT.
The increase in the initial stage observed in dose 0% may be
linked to intense multiplication of microorganisms, resulting in a
marked increase in turbidity, since the microorganisms are also
particulate or suspended biomass (Jero ^nimo et al., 2012).
After the consumption of most of the substrate (organic matter
in swine and glycerin), microorganisms begin to use their proto-
plasmic reserves for survival, i.e., catabolize its cellular reserves
(cell decline/death phase, as exhibited in Fig. 3b), and the popula-
tion of microorganisms tends to decrease. It is in the endogenous
phase (cell death) that the formation of biological flakes occurs,
with subsequent decantation, resulting in a decrease in turbidity,
which can be observed at dose 0% after 35 days of HRT, much like
the theoretical behavior shown in Fig. 3b (Jero ^nimo et al., 2012).
Thus, the process of microbial catabolism promotes the degra-
dation of assimilated organic compounds in final products such as
CO2, CH4 and H2S (Miki, 2010), promoting the reduction of
turbidity.
For the treatments with glycerin doses, a decrease on the initial
values of turbidity (4170 NTU) was observed, in a first moment this
is due the dilution effect, considering that the added glycerin was
translucent, diluting the initial turbidity of the swine manure. In a
second moment, the microorganisms inside the reactors adapted to
120
100
Ntotal (gkg-1)
80
60
40
20
0 0.0
7 14 21
HRT (d)
(P) Dose 0% (P) Dose 5%
(N) Dose 0% (N) Dose 5%
Fig. 4. Average Ntotal and Ptotal concentrations in the biodigesters in function of hydraulic retention time (HRT) during 42 days of evaluation. Note: Concentrations of Ntotal and Ptotal
of swine manure in HRT 0 days: 79.62 ± 12.30 and 1.14 ± 0.30 gkg-1, respectively.
179
the new condition of substrate. Thus, it is observed that some of the
doses of glycerin were high and possibly impaired the initial
development of the microorganisms, because only after 35 days it is
observed an increase of the turbidity of the medium.
A reduction of Ntotal concentrations in the treatments with
glycerin addition is observed in Fig. 4. This occurs probably due to
the high carbon concentration in glycerin, since its molecule
(propane-1,2,3-triol) is composed of carbon (C), hydrogen (H) and
oxygen (O) atoms.
Another justification for the reduction of Ntotal is due to the
removal of N in volatile forms, such as NH3, one of the biogas
components (Wiesmann et al., 2007; Vivan et al., 2010).
According to Costa et al. (2016), the low N removal rate may be
associated with acidification of the effluent within the bioreactor,
which may have compromised the degradation of more complex
compounds such as macronutrients, which vary in swine manure
according to age, race, diet composition, water consumption of the
animals and environmental factors.
The addition of glycerin in biodigestors causes a dilution on
Ntotal concentration in the medium. Also, the higher carbon content
may increase the development of the anaerobic microorganisms
until certain point, accelerating even more the microbial absorption
of N and volatilization in the form of NH3.
As glycerin contains a high carbon content of easy assimilation,
it can be considered that as a co-product of an anaerobic digestion it
enriches the process, since carbons in more complex structures,
such as long chain fatty acids, can become toxic. Therefore glycerol
is indicated for co-digestion with animal wastes having N to bal-
ance the medium (Schwingel et al., 2017).
According to Fig. 4, there were increases in Ptotal concentrations
along the HRT in the evaluated treatments. This result is possibly
due to the biodigestion process, with the occurrence of decompo-
sition of the waste, releasing organic P from the solid phase of the
waste to the liquid phase, which is measured in the form of Ptotal.
The main problems caused by swine to the environment are
pollution of soil and water resources, mainly due to the waste
physical and chemical composition. Maggi et al. (2013) found that
the levels of N, P and K in the soil increased considerably by the
increase of application rates of swine manure.
Schwantes et al. (2017b), studying the anaerobic digestion of
organic residues from vegetable origin aiming the production of
biofertilizer, found concentrations of P ranging from 1.0 to 1.8
2.5
2.0
Ptotal (gKg-1)
1.5
1.0
0.5
28 35 42
(P) Dose 10% (P) Dose 15% (P) Dose 20%
(N) Dose 10% (N) Dose 15% (N) Dose 20%
180 M.L. Veroneze et al. / Journal of Cleaner Production 213 (2019) 176e184

mgL
1, considered by the authors as insufficient for plant nutrition

purposes.
In this research, the mean concentration of Ntotal and Ptotal
exhibited a decrease tendency due to the addition of increasing
doses of glycerin (Table 2), possibly due to the action of two
different processes, first the dilution caused by the addition of
glycerin, and second the acceleration of biodigestion process, due to
the greater availability of substrate to the microorganisms, which
consume these nutrients in the liquid medium.
The average contents of N and P in the co-digested wastes (20.09
gNL
1 and 0.83 gPL
1) are higher than those contained in the
biodigestion of vinasse studied by Silva et al. (2016) (0.71 gNL
1 and
0.17 gPL
1), demonstrating that swine manure co-digested with
glycerin may represent an excellent source of these nutrients for
plants.
Schwantes et al. (2017a) evaluated the use of biodigested sludge
from the sewage treatment plant of the whey industry as fertilizer
for tifton 85 grass, obtaining Ntotal and Ptotal contents of 6.53 and
0.98 gkg-1, again, N levels lower than those found in this study.
Chiconato et al. (2013) evaluated the use of biostabilized bovine
manure in lettuce fertilization, and observed that after the bio-
digestion process, N and P contents were respectively 0.14 and
0.054 mgL
1, lower than this research, indicating that swine
manure may represent a good source for N and P in vegetable
fertilization, even after glycerin co-digestion.
3.2. Concentration of K, Ca, Mg, Cu, Zn and Mn
Fig. 5 shows the mean total values of K, Ca, and Mg concentra-
tions in the liquid phase of swine manure and glycerin mixture
during the 42 days of HRT.
The metals Ca, Mg and K are constituents of the FS fraction, i.e.,
these elements are not transformed into any component of the
biogas. The fluctuation of its concentrations in the liquid medium
possibly occurs as a function of the medium pH variations, as a
function of the acid and neutral pH phases of the biodigestion, with
consequent formation of soluble or insoluble complexes, or by the
decomposition of the solid particles of swine manure, which
gradually makes such elements available and solubilizes them in
the reactor as the biodigestion proceeds.
Astals et al. (2012, 2013) did not observe significant variation of
K contents after the anaerobic treatment of swine manure with
glycerin, demonstrating that the addition of glycerin doses appar-
ently does not cause significant reductions on K contents in the
liquid medium.
Fig. 5. Average total concentrations of potassium (K) calcium (Ca), magnesium (Mg) in
the biodigesters during 42 days of HRT. Note: Concentrations of K, Ca and Mg of swine
manure in HRT of 0 days: 256.70 ± 95.02; 14.00 ± 2.20; 10.60 ± 18 mgL
1, respectively.
The mean concentrations in the present study were 0.77, 0.097
and 0.073 mgL
1, for K, Ca and Mg, respectively, these concentra-
tions are higher than those found by Aguiar et al. (2017) when using
bovine biofertilizer in yellow passion fruit fertilization, whose
concentration of K, Ca and Mg were, respectively, 0.023, 0.013,
0.049 gL-1. The same authors report a rise in fruit production per
plant and, consequently, productivity, by using this biofertilizer.
Chiconato et al. (2013), investigated the use of biostabilized
bovine manure and its use in biofertilization of lettuce, with
average levels of 0.048, 0.065 and 0.026 gL-1 respectively for K, Ca
and Mg, (Table 2), i.e., the swine manure þ glycerin exhibited
average levels of K, Ca and Mg, 15.4, 1.49 and 2.8 times higher than
the obtained by Chiconato et al. (2013).
It is worth mentioning that Chiconato et al. (2013) found an
increase in the components height, number of leaves, diameter and
fresh mass of lettuce plants with the application of 60 m3ha-1,
evidencing that the application of biofertilizers can be a sustainable
practice.
A variation is observed of metal contents in the liquid medium
of the wastes during biodigestion process. This is due to the
biochemical processes that occur during biodigestion, being influ-
enced by the pH, temperature, availability of nutrients to micro-
organisms, among other factors that may be related to the
availability of these elements to the liquid medium.
Chiconato et al. (2013) used biostabilized bovine manure in
organic fertilization of lettuce and obtained Cu, Zn, Fe and Mn

Table 2
Total contents of N, P, K, Ca, Mg, Cu, Zn, Fe and Mn in swine waste þ glycerin and comparison with other organic residues cited by literature.

Organic waste N P K Ca Mg Cu Zn Fe Mn Reference

gL
1 mgL
1

Swine waste þ glycerin (HRT ¼ 42 days Dose 0%) 38.50 1.77 1.39 0.090 0.039 0.24 3.65 1.15 0.82 Present research
Swine waste þ glycerin (HRT ¼ 42 days Dose 5%) 26.77 1.03 0.99 0.097 0.110 0.45 1.61 3.34 6.75 Present research
Swine waste þ glycerin (HRT ¼ 42 days Dose 10%) 15.75 0.51 0.76 0.094 0.011 0.15 4.00 3.47 4.83 Present research
Swine waste þ glycerin (HRT ¼ 42 days Dose 15%) 9.10 0.54 0.72 0.12 0.108 0.26 4.10 4.28 4.69 Present research
Swine waste þ glycerin (HRT ¼ 42 days Dose 20%) 10.32 0.31 0.74 0.14 0.127 0.30 4.10 4.62 4.07 Present research

Mean values obtained in the present study (42 days) 20.09 0.83 0.77 0.097 0.073 0.28 3.48 3.37 4.23 This research

Swine waste 19.60 23.60 159.10 27.70 12.20 30.00 117.00 303.00 110.00
coli et al. (2016)
Bo
Bovine biofertilizer 0.14 0.05 0.05 0.065 0.026 0.10 1.40 1.00 0.20 Chiconado et al. (2013)
STS sludge, whey industry 6.53 0.98 0.78 1.89 1.29 5.34 15.00 440.00 13.85 Schwantes et al. (2017)
Vinasse 0.71 0.17 Silva et al. (2016)
Bovine biofertilizer 0.06 4.33 6.77 8.50 Ferreira et al. (2015)
Cattle manure 0.78 0.73 1.19 0.590 0.28 2.04 5.90 73.04 9.32 Alencar et al. (2015)
Wastes, urine and dietary remains of dairy cows 4.25 0.02 0.13 0.30 0.37 Soares Filho et al. (2015)

Transformed values for gL
1 and mgL
1, for comparative purposes.

 M.L. Veroneze et al. / Journal of Cleaner Production 213 (2019) 176e184 181

contents of 0.1, 1.4, 1.0, 0.2 mgL
1. At 42 days of HRT, the swine
wastes þ glycerin exhibited concentrations of 0.28, 3.48, 3.37 and
4.52 mgL
1 for Cu, Zn, Fe and Mn respectively, i.e., concentrations
2.8, 2.48, 3.37 and 22.6 times higher than the bovine manure
studied by Chiconato et al. (2013) (Table 2).
This result (Fig. 6) show that micronutrient concentrations are
quite significant for plant fertilization, and the biofertilizer evalu-
ated is a good alternative for the supply of Cu, Zn, Fe and Mn for
plants.
Toxic metals such as Cr and Cd were not detected in evaluations,
however, concentrations of Pb not higher than 0.2 mgL
1 were
found in two single determinations. Pb may be in swine manure
due to the contamination of mineral supplements. Unlike Cu, Zn
and Mn, toxic metals such as Pb are not essential to plant or animal
growth and can be toxic even at low concentrations (Moral et al.,
2008).
Moreover, the presence of harmful substances in the substrate,
such as toxic metals can cause adverse effects to the bacteria,
inhibiting the fermentation process (Wohlgemut et al., 2011).
3.3. Concentrations of TS, FS, VS and COD
Higher glycerin doses result in higher TS concentration (Fig. 7).
This occurs because adding glycerin in the system, the increase the
concentration of SV take place, since glycerin [C3H5(OH)3] is basi-
cally composed by carbon, hydrogen and oxygen atoms, favoring
the production of biogas components, mainly CH4, CO2, NH3 and
H2S. The concentration of solids refers to the total residue in the
substrate, whether organic or inorganic, and is an indicator of the
total mass to be treated (Leite and Povinelli, 1999).
Schwingel et al. (2016) studied the co-digestion of swine excreta
associated with increasing doses of crude glycerin and obtained the
best reductions of TS with HRT of 24 days, obtaining reductions of
40%, 54.9%, 31.1% and 49% with doses of 0, 5, 10 and 15%,
respectively.
Konrad et al. (2014), by adding 6% glycerol in manure observed
reductions for TS concentration, exhibiting pronounced results in
control treatment, with 28% reduction, and treatment with 9% of
glycerin, 15% reduction of TS.
The highest concentrations for VS are obtained at the doses of
15% and 20% glycerol (Fig. 7), a result that was already expected,
since the higher the glycerin addition the greater the amount of
volatile compounds in the mixture.
VS are the main contributors to biogas production. The higher
the VS concentration in the daily feed of the biodigester, the greater
is the biogas production and, consequently, the greater the energy
production (Lima, 2007). The VS are basically composed of organic
matter. They comprise the carbonaceous material that will be
biodigested and transformed into biogas (Costa et al., 2016).
For Elango et al. (2007), the VS content is an important
parameter to measure biodegradation, since it indicates the
metabolic state of some of the most delicate groups of microor-
ganisms in the anaerobic system, so the reduction of SV indicates
the process stabilization.
Regarding COD (Fig. 7), the treatment with 20% glycerin had
higher COD values, followed by 15%, 10%, 5% and 0%. Similar to SV,
the higher the amount of added glycerin, the higher the COD
concentrations.
COD is an important parameter to be monitored during anaer-
obic digestion, since it indicates the stabilization of organic matter.
By knowing the initial COD values of the manure and the COD
concentration after treatment, it is possible to calculate the COD
removal efficiency, and the higher the efficiency, the more biode-
gradable and the biostabilized is the final residue (Sgorlon et al.,
2011).
It is observed that the control treatment remained constant in
relation to HRT of 0 days. The COD variation was low among the
doses. The dose 20% exhibited the highest COD concentrations. This
can be explained by the occurrence of a possible overload of the
glycerin in the reactor, which can cause adsorption and drag of
solids to the bottom, interfering in the removal of COD.
The low COD removal along the HRT is credited to the difficulty
of anaerobic microorganisms in effectively degrading the mixture
of swine manure with glycerin at studied doses and in reactors
without stirring.
In a non-stirred reactor, the substrate tends to form layers due to

13.0 5.5
12.0 5.0
11.0

Cu and Zn (mgL-1)
4.5
Fe and Mn (mgL-1)

10.0
4.0
9.0
8.0 3.5
7.0 3.0
6.0 2.5
5.0 2.0
4.0
1.5
3.0
1.0
2.0
1.0 0.5
0.0 0.0
7 14 21 28 35 42
HRT (d)
(Cu) Dose 0% (Cu) Dose 5% (Cu) Dose 10% (Cu) Dose 15% (Cu) Dose 20%
(Zn) Dose 0% (Zn) Dose 5% (Zn) Dose 10% (Zn) Dose 15% (Zn) Dose 20%
(Fe) Dose 0% (Fe) Dose 5% (Fe) Dose 10% (Fe) Dose 15% (Fe) Dose 20%
(Mn) Dose 0% (Mn) Dose 5% (Mn) Dose 10% (Mn) Dose 15% (Mn) Dose 20%

Fig. 6. Average total concentrations of copper (Cu), zinc (Zn), iron (Fe) and manganese (Mn) in the biodigesters during 42 days of HRT. Note: Average concentrations of Cu, Zn, Fe and
Mn of swine manure in HRT at 0 days: 0.30 ± 0.06; 0.89 ± 0.24; 0.47 ± 0.20 and 1.67 ± 0.34 mgL
1, respectively.
182 M.L. Veroneze et al. / Journal of Cleaner Production 213 (2019) 176e184

350 2400

300 2100
TS and VS (mg.L-1)

1800
250

COD (mg.L-1)
1500
200
1200
150
900
100
600
50 300
0 0
7 14 21 28 35 42
HRT (d)
(COD) Dose 0% (COD) Dose 5% (COD) Dose 10% (COD) Dose 15% (COD) Dose 20%
(TS) Dose 0% (TS) Dose 5% (TS) Dose 10% (TS) Dose 15% (TS) Dose 20%
(VS) Dose 0% (VS) Dose 5% (VS) Dose 10% (VS) Dose 15% (VS) Dose 20%

Fig. 7. Average concentration of TS, VS and COD (mgL
1) during 42 days of anaerobic biodigestion of swine manure and doses of glycerin (0, 5, 10, 15 and 20%). Note: Concentrations
of TS, VS and COD of swine manure in TDH of 0 days: 17.57 ± 0.33; 13.33 ± 0.34 and 655.67 ± 100.20 mgL
1, respectively.

the differences in density found in the waste. A large part of the
substrate tends to occupy the lower parts, limiting the processes of
biodegradation of the substrate, thus causing the drag of solids to
the bottom of the biodigesters.
The process of biodigestion consists of optimizing the degra-
dation of the organic matter of the waste, allowing the reduction of
COD, BOD and VS, making the nutrients in forms more available for
plant absorption (Orrico Júnior et al., 2010).
3.4. Biogas production
It was observed that the dose without addition of glycerin had
the highest rate of biogas production at the start of biodigestion
process, reaching 1.67 LL
1d
1 at 42 days of HRT, with the dose of
5% in second place, with 1.16 LL
1d
1, and the other doses, with
lower rates of biogas production as glycerin doses increased (Fig. 8).
It is noteworthy that only in Brazil, during 2016, glycerin pro-
duction reached 430,807 kg (equivalent to 341.9 m3) (ANP, 2016),
which may not be used in the various industrial segments in an
integral way. This glycerin, when added in biodigesters containing
swine manure (5% vv
1 ratio) can promote the biogas production
capacity up to 1.16 LL
1d
1.
Apparently, biogas production rates were adversely affected by
the addition of glycerin, this corroborates with the results from
Fig. 3 (turbidity), where the addition of glycerin caused a slower
adaptation of microorganisms to the medium, and in those cases,
the biodigestion evaluated period was short for more efficient
biogas production.
The high C/N rate causes concern due to the system overload,
being motivated by the inhibition of the methanogenic activity,
depending on the reactor conditions and the degree of adaptation
of the microbial community (Rajagopal et al., 2013; Vrieze et al.,
2015).
According to Fig. 9, a quadratic relation between the concen-
tration of VS and the biogas production is found for the control
treatment and dose 5%.

1.80
1.60
Biogas production rate

1.40
1.20
(LL-1d-1)

1.00
0.80
0.60
0.40
0.20
0.00
0 5 10 15 20
Glycerin doeses (%)
7 days 14 days 21 days 28 days 35 days 42 days

Fig. 8. Biogas production rate (LL
1d
1) during hydraulic retention time (HRT).
 M.L. Veroneze et al. / Journal of Cleaner Production 213 (2019) 176e184
1.8
Biogas production rate (LL-1d-1)
Biogas= 0.001[VS]2 - 0.1021[VS] + 2.8353
1.6
1.4
1.2
1.0
0.8
0.6
0.4
0.2
0.0
10 20
Fig. 9. Relation between biogas production rate (LL
1d
1) and VS during hydraulic retention time (HRT) for the addition of 0% and 5% glycerin.
Using crude glycerin in co-digestion with swine manure, in
semi-continuous biodigesters with mesophilic regime, Astals et al.
(2012) verified an increase of about 400% in biogas production with
4% glycerol (mm
1). The authors correlated this result with the
increase of organic load and the optimum C/N rate, which was
complementary between the two waste materials.
Amon et al. (2006) commented that for a stable process in
biodigestion, the amount of added glycerin should have a limiting
concentration level. In their studies the added crude glycerin
should not exceed 6%.
Holm-Nielsen et al. (2008), studying manure and food industry
residues with glycerin at concentrations of 0e6.5%, reported that
the addition of glycerin above this level may cause an organic
overload, and the rate of methane production declines, and may
even be interrupted.
Fountoulakis et al. (2010) also demonstrated that crude glycerin
may be advantageous in biodigestion, but a strict control of the
added amount should be avoided, reducing the risk of organic
overload in the biodigester.
The same authors pointed out that this inhibitory tendency in
the production of biogas with high concentrations of crude glycerin
may be associated to the contamination of glycerin by biodiesel
residues or an organic overload in the biodigester.
It is important to point out that the state of Parana
is notably
known as a large swine producer. According to estimates, only in
the year 2017, 9,203,619 pigs were slaughtered in this state (Seab,
2018). Considering that on average a pig produces 8.60 Ld-1
(SEMA 031, 1998), it is estimated the production of 28,890,160 m3 of
swine manure in the state of Parana
only in 2017, i.e., a huge
amount of manure with enormous pollution potential.
Considering the maximum biogas production rate obtained in
this research (1.67 LLd
1), it is estimated that only in the State of
Paran
a, it is possible to produce 17,299,497 m3 of biogas per year, or,
considering that 1 m3 of biogas can produce around 1,25 KWh,
about 21,624 MWh of electricity.
Much of this waste is destined to agriculture without any
treatment, i.e., without any type of biodigestion, which has caused
numerous pollution problems to the soil, surface water and
groundwater.
The results shown in this research demonstrate that this enor-
mous amount of residues can be treated by means of co-digestion
with glycerin, when applied at doses lower than 5% vv
1, possibly
increasing the biogas production and potentiating the use of the
183
Dose of 0% Dose 0%
R² = 0.7919 Dose 5%
Dose of 5%
Biogas = 0.0071[VS]2 - 0.8328[VS] + 24.416
R² = 0.9563
30 40 50 60 70 80
VS (mgL-1)
final residue as biofertilizer in agriculture, with lower environ-
mental risks to soil and water.
4. Conclusions
Anaerobic co-digestion of swine manure and glycerin didn't
promoted higher production of biogas when compared with the
digestion without addition of glycerin. When the amount of crude
glycerin exceeded 5% a tendency was observed for the reduction of
the biogas production according to the increases in the glycerin
amounts.
Very high doses of glycerine, such as 20%, harmed the produc-
tion of biogas, possibly because of the elevation of C/N relation. It's
important to emphasize that this relation is essential to the mi-
crobial development and for the stabilization of the biofertilizer.
Apparently, biogas production rates were impaired by the
addition of glycerin because in reactors with glycerin addition the
adaptation of the microorganisms to the reaction medium was slow
and the biodigestion period evaluated was short.
Because of the accumulation of crude glycerin in biodigesters, it
is believed that the use of stirring systems would reduce this type of
problem and would probably allow a better evaluation of the in-
fluence of the residue on the process.
The obtained results demonstrate that nutrient concentrations
in the biostabilized waste are expressive for vegetal fertilization, so
it can be used for this goal.
Acknowledgments
~o Arauc

This research was supported by the Fundaça aria.
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