XXXIII IULTCS Congress

November, 24th 27th, 2015 Novo Hamburgo/Brazil

158

Biogas production from tannery wastes Evaluation of isolated

microorganisms effect

Jayna Pessuto, Marcelo Godinho, Aline Dettmer

1
Engineering of Processes and Technologies Post-Graduate Program - University of Caxias do Sul - Francisco
Getlio Vargas Street, 1130, ZIP CODE: 95070-560, Caxias do Sul, RS, Brazil. Phone: +55 54 3218.2100

Abstract
The disposal and treatment of wastes generated in the leather industry has been the focus of several
studies around the world. They intend to meet environmental legislation and make the process
environmentally and economically viable. The anaerobic digestion process, which occurs in steps that
involve acid and methane forming bacteria, can be an alternative since it turns the waste into
renewable energy. This work evaluated the anaerobic digestion process and the generation of biogas
from leather industry waste (wet-blues shavings and sludge from wastewater treatment plant - WWTP)
and from parboiled rice waste. In both, microorganisms isolated from aerobic sludge of WWTP
(inoculum) were added. The waste was analyzed for organic carbon and total solids content. The
samples were maintained at 35C, the volume of gas generated was measured and the composition was
determined by gas chromatography. The addition of inoculum increased volume of biogas generated.
The amount of biogas generated by leather waste was 23 ml (composed of 25% of hydrogen, in molar
basis).The rice with the addition of tannery WWTP sludge generated 47 ml of biogas (composed of
75% of hydrogen, in molar basis). The anaerobic digestion process is efficient for leather industry
waste treatment since it converts waste into renewable energy.

Keywords: anaerobic digestion, waste leather, wet-blue, inoculum, biogas.

1 Introduction

The treatment and the correct disposal of waste is a global concern. Its lack may generate numerous
consequences for the environment and for the society. It is estimated that the Brazilian leather industry
generates more than 202,000 tons of leather waste tanned with chromium per year.

Anaerobic digestion can be a promising alternative for the treatment of this waste since it converts
waste into energy (biogas). Biogas is composed of methane (60%), carbon dioxide (40%), and trace
amounts of water vapor, hydrogen sulphide and ammonia. It is an odorless and colorless gas that
produces a blue flame when burned, similar to the one of liquefied petroleum gas (CHRISTY et al.,
2014). Among the main applications of biogas, it can be mentioned: heat production, steam production
and combined electricity and heat production (ABBASI et al., 2012 and JEIHANIPOUR et al., 2013).

The conversion of complex organic matter to methane by anaerobic digestion (AD) process requires a
consortium of microorganisms. It comprises several groups of anaerobic microorganisms that interact
in a metabolic manner (COVINGTON et al., 2003).

The substrates most often employed for waste conversion into energy are manure (54%), sewage
sludge (22%) and municipal solid waste (11%). In order to improve the AD process , other residues
are added to the process, being the industrial waste (41%), agricultural wastes (23%) and municipal

1
 XXXIII IULTCS Congress
November, 24th 27th, 2015 Novo Hamburgo/Brazil

waste (20%) the most used ones (ALVAREZ et al., 2014). Co-digestion is defined as the anaerobic
treatment of a mixture of at least two different substrates in order to increase the efficiency of the AD
process. It makes possible to adjust the C/N ratio (KAFLE & KIM, 2013).

Anaerobic digestion can be divided into four main phases: hydrolysis, acidogenesis, acetogenesis and
methanogenesis. In the first stage (hydrolysis), large molecules of protein, fat and carbohydrate
polymers (such as cellulose and starch) are hydrolyzed in water-soluble monomers (amino acids, long
chain fatty acids and sugars). Hydrolysis is caused by extracellular enzymes (hydrolases) such as
lipases, proteases, cellulases (LI et al., 2011 and MOLINO et al. 2012). In the second stage, the
acidogenic bacteria produce intracellular enzymes that convert the hydrolysis products in volatile fatty
acids and alcohol. In the third stage, acetogenic bacteria (or homoacetogenics) convert the product of
simple organic phase into acids, carbon dioxide and hydrogen, occurring the production of acetic acid,
butyric acid, propionic acid and ethanol. In the fourth stage, the methane can be produced by two
routes: a) from acetate - conversion of methyl and carboxyl groups of acetate to methane and carbon
dioxide by the action of methanogenic bacteria (process known as direct methanogenesis) and b) by
oxidation of the acetate to carbon dioxide and reduction of the methane. The latter route occurs
through oxidizing and methanogenic hydrogenotrophic bacteria (SASAKI et al., 2011; MOLINO et al.
2012; ABBASI et al. 2012 and COVINGTON et al. 2003).

Factors such as pH, temperature and total solids content directly influence the AD process and its
products. Low temperatures (12 to 16C) can result in the exhaustion of microbial cell energy. In
contrast, high temperatures (above 40C) lead to low yield of methane in the gas generated due to
production of volatile gases, such as ammonia, which suppresses the methanogenic activity. The
optimum pH for the methanogenic bacteria was set around 7.0. There are reports that the hydrolysis
and acidogenesis occurs at pH values between 5.5 and 6.5. The total solids content of the substrate
influences the AD process. Tests conducted with total solids content progressively varying from 10 to
35% reduced the methane production (ABASSI-GUENDOUZ et al., 2012). The efficiency of the AD
process can be affected by pretreatments. They aim to increase the kinetics of the process wastes AD
and are responsible for an increase on biogas production efficiency. Ultrasound is one of the most
commonly used pre-treatments (33%), followed by thermal pretreatment (24%) chemical
pretreatments (21%) (ALVAREZ et al., 2012). The chemical pretreatment conducted in the waste of
leather tanned with chromium salts and vegetable tannins provided a higher yield in biogas production
when compared to the samples without pretreatment (DHAYALAN et al., 2007).

The objective of this study was to evaluate the influence of inoculum addition in the leather industry
waste for biogas generation

2 Materials

Parboiled rice waste was provided by Caxiense Company of Biological Control Ltda (CCBC). It was
obtained from a culture of the fungus Trichoderma spp., which is used for preventive and curative
control of diseases caused by phytopathogenic fungi in plants. The tannery sludge (TLS) was obtained
from a local tannery wastewater treatment plant (WWTP). The same company provided chromium
tanned leather shavings. The sludge from the University of Caxias do Sul (UCS) WWTP was made
available by the institution.

2
 XXXIII IULTCS Congress
November, 24th 27th, 2015 Novo Hamburgo/Brazil

3 Methods

3.1. Waste characterization

The determination of the percentage of moisture and total solids content (TS) of rice and leather
wastes were based on the method ASTM D3790-12. The total solids content of WWTP sludge
samples were analyzed based on the standard ABNT NBR 14550. The organic carbon content was
determined by means of the sample carbon oxidation in acidic medium followed by titration with
ferrous sulphate. The methodology was based on Walkley-Black (1934).

3.2. Isolation and identification of microorganisms

Isolation and identification of microorganisms are described in the work of Silva (2015). The access
permission to genetic resources was given by the Brazilian Institute of the Environment (IBAMA),
with authorization number 02001.003486/2013-86.

3.3. Inoculum

The microorganisms previously isolated (five in total) were used to prepare the inoculum. Aliquots of
isolates were added to the LB culture medium (liquid medium - LM) and placed in a thermostatic bath
at 35C until medium optical density (OD), measured at 600 nm, equal to 1.0.

3.4. Anaerobic Digestion Assay

The influence of the addition of the inoculums was evaluated through the volume and molar gas
fraction. The gas samples were generated by leather pretreated with chemical and thermal process and
by tannery sludge added to rice waste. The thermal treatment of leather waste consisted of placing it in
an airtight glass container and autoclaving it (in a Prismatec, model CS, equipment) at a pressure of
1atm for 15 minutes. The chemical pretreatment method was adapted from the methodology used by
Dhayalan et al. (2007), using 200% distilled water, 0.003 g of oxalic acid and 0.003 g of
ethylenediamine tetra acetic acid (ETAA).

The samples were placed in flasks with a total volume of 100 mL. The test was conducted at 35C for
as long as biogas was being generated. Inoculum or LM was added after 24 hours of testing and then
every 7 days, totalizing five additions. The volume of inoculum or LM was added such that the final
TS content did not exceed 9%. After the tests, the reduction of organic carbon and TS was evaluated.
Table 1 lists the samples evaluated in the AD process.

3
 XXXIII IULTCS Congress
November, 24th 27th, 2015 Novo Hamburgo/Brazil

Table 1. Description of the samples analyzed during AD process.

3.5. Volume and molar fraction of generated biogas

The volume and the molar fraction of the biogas generated in each sample were measured daily. The
verification of the generated gas volume was performed using the volumetric method. For the
chromatographic analysis, nitrogen was used as carrier gas. Thermal conductivity detector and
capillary column Supelco Carboxen TM 1006 (30m x 0,53mm) were employed. For sample collection,
it was used a Gasthight 1 ml Hamilton syringe (SILVA, 2015).

4 Results and discussion

4.1. Waste characterization

Table 2 shows the results of organic carbon, chromium, TS and moisture content of the waste used
during testing on a dry basis.

Table 2. Content of organic carbon, total solids, moisture and chromium in waste

4.2. Isolation of microorganisms

Five isolations were performed at different times of testing; the identification showed that the
microorganisms belong to the genus Bacillus.

4.3. Anaerobic digestion assay with addition of microorganisms

Figure 2 compares the influence of the addition of inoculum and LM to the leather waste chemically
and thermally pretreated on the volume and molar fraction of the biogas generated.

4
 XXXIII IULTCS Congress
November, 24th 27th, 2015 Novo Hamburgo/Brazil

5
 XXXIII IULTCS Congress
November, 24th 27th, 2015 Novo Hamburgo/Brazil

Figure 2. Influence of the addition of LM or inoculum on the volume and molar fraction of gas
generated from pretreated leather waste.

It may be observed that the leather waste with addition of inoculum generated larger volume of biogas
when compared to the samples with the addition of LM and to the ones of leather waste without
addition of inoculum. The gas generated was predominantly composed of carbon dioxide. The molar
fraction of hydrogen in the samples with addition of LM (0.10) decreased when compared to the
samples with addition of inoculum (0.24). This result confirms the importance of the presence of
microorganisms that assist in the AD process.

The predominant mole fraction of carbon dioxide in the gas produced from leather waste and its
combinations causes a contamination in the biogas, reducing its calorific value. However, the addition

6
 XXXIII IULTCS Congress
November, 24th 27th, 2015 Novo Hamburgo/Brazil

of CO2 in digesters to treat food waste and sludge from municipal WWTP was evaluated by
FRNANDES et al (2014). The authors added different partial pressures of CO2 in a reactor. They
noticed an increase in 13% for methane production and consequent carbon dioxide reduction on the
gas obtained, ranging from 8 to 34% for sludge of municipal WWTP. According to the authors, with
CO2 enrichment, the acetoclastic microbial activity was increased, which resulted in an increase in the
molar fraction of methane (FRNANDES et al., 2014).

Figure 3 shows the mole fraction of gas generated from the addition of LM or inoculum in rice waste
samples with tannery sludge.

7
 XXXIII IULTCS Congress
November, 24th 27th, 2015 Novo Hamburgo/Brazil

Figure 3. Volume and molar fraction of biogas generated from the tannery sludge waste added to the
rice waste

It can be seen from the results of Figure 3 that the sample containing rice and tannery sludge presented
similar gas volumes and hydrogen mole fraction close to 1.0, independently of the addition of
inoculum or LM.

Acetogenic bacteria typically produce hydrogen and, for each four moles of hydrogen consumed by
methanogenic hydrogenotrophic microorganisms, one mole of carbon dioxide is converted to methane
(ZHANG et al., 2014).

8
 XXXIII IULTCS Congress
November, 24th 27th, 2015 Novo Hamburgo/Brazil

The fermentation process produces hydrogen from the decomposition of carbohydrates present in
waste. Polysaccharides, such as starch, are hydrolyzed and converted into low molecular weight
compounds that are transported into the interior of the bacterial cells and converted into products of
interest, such as hydrogen (TREVISAN, 2010).

The step of anaerobic digestion that produces hydrogen is enabled by acidogenic microorganisms,
which decompose the substrates in hydrogen, carbon dioxide and volatile fatty acids. The reaction is
represented by the equations below (PEIXOTO, 2011):

The reduction of organic carbon content and solids content of the waste used in this experiment is
presented in Table 4.

Table 4. Content of organic carbon, total solids content and percentage reduction after AD process

It is possible to affirm, based on Table 4, that the carbon contents and TS, after the process, reduced.
The rice waste with tannery sludge and inoculum addition showed greater reduction in carbon
(77.88%) and in total solids content (90.34%).

It indicates an effective AD process of the wastes studied in this work and show the importance of
inoculum addition to improve the process.

5 Conclusions

The addition of the inoculum to wastes of leather chemically and thermally pretreated resulted in an
increase in the generated gas volume. The molar fraction of hydrogen generated in samples with added
inoculum was higher when compared the samples with addition of LM. The predominant molar
fraction of the biogas generated from the addition of tannery sludge to rice waste was hydrogen
(approximately 1.0) independently of the addition of inoculum. The reduction in organic carbon and
TS showed an effective anaerobic digestion process in the tests performed in this study.

6 Acknowledgements

The authors thank to FAPERGS and CNPQ for the financial support of this study.

7 References

1. AMERICAN SOCIETY FOR TESTING AND MATERIALS. ASTM D3790: Standard Test
Method for Volatile Matter (Moisture) of Leather by Oven Drying. West Conshohocken:
2012.

9
 XXXIII IULTCS Congress
November, 24th 27th, 2015 Novo Hamburgo/Brazil

2. BRAZILIAN ASSOCIATION OF TECHNICAL STANDARDS. ABNT 14550. Leather -

Bath and residual liquid effluent - Determination of solids content of total dissolved, fixed and
volatile. Brazil, 2015.
3. ABBASI, T.; Tauseef, S.M.; Abbasi S.A (2012) Anaerobic digestion for global warming
control and energy generationAn overview. Renewable and Sustainable Energy
Reviews16:3228 3242.
4. ABASSI-GUENDOUZ, A. etal.Total solids content drives high solid anaerobic digestion via
mass transfer limitation. Bioresource Technology, 111, 55-61, 2012.
5. COVINGTON, A.D. et al. Biodegradation of solid leather wastes. MaterieConcianti, 79, 187-
195, 2003.
6. CHIRSTY, P.M. et al. A review on anaerobic decomposition and enhancement of biogas
production through enzymes and microorganisms. Renewable and Sustainable Energy
Review, 34, 167-173, 2014.
7. DHAYALAN, K. et al.Biodegradability of leathers through anaerobic pathway. Waste
Management,27, 760- 767, 2007.
8. JANG, H. et al. Influence of thermophilic aerobic digestion as a sludge pre-treatment and
solids retention time of mesophilic anaerobic digestion on the methane production, sludge
digestion and microbial communities in a sequential digestion process. Water Research,48, 1-
14, 2014.
9. JEIHANIPOUR, A. et al. High-rate biogas production from waste textiles using a two-stage
process. Renewable Energy, 52, 128-135, 2013.
10. KALID, A. et al. The anaerobic digestion of solid organic waste. Waste Management, 31,
17371744, 2011.
11. KAFLE, B. et al. Anaerobic treatment of apple waste with swine manure for biogas
production: Batch and continuous operation. Applied Energy,103, 61-72, 2013.
12. KILI, E.et al. Chromium recovery from tannery sludge with saponin and oxidative
remediation.Journal of Hazardous Materials, 185, 456-462, 2011.
13. LI, Y.; PARK, S. Y.; ZHU, J. Solid-state anaerobic digestion for methane production from
organic waste. RenewableandSustainable Energy Reviews, 15, 821-826, 2011.
14. MAO et al. Review on research achievements of biogas from anaerobic digestion. Renewable
and Sustainable Energy Reviews, 45, 540555, 2015.
15. MATA-ALVAREZ, J. et al.A critical review on anaerobic co-digestion achievements between
2010 and 2013. Renewable and Sustainable Energy Reviews,36, 412-427, 2014.
16. MOLINO A. et al.Biomethane production by anaerobic digestion of organic waste. Fuel, em
impresso, 2012. Disponvel em: . Acessoem: 20 mar.2013, 13:30.
17. PEIXOTO, G. Two Phase fermentation system for Hydrogen Production and Methane from
Sewage Combined with Health Industrial Wastewater.2011,214 f. Doctoral thesis. (Graduate
Engineering) Universityof So Carlos, So Paulo, 2011.
18. SHANMUGAM, P & HORAN, N.J. Optimising the biogas production from leather fleshing
waste by codigestion with MSW. Bioresource Technology, 100,4147-4120, 2009.
19. SASAKI, D. et al. Methanogenic pathway and community structure in a thermophilic
anaerobic digestion processo of organic solid waste. JournalofBioscienseandBioengineering,
1, 41-46, 2011.
20. SILVA, P. J. Biogas generation from co-digestion of waste agroindustrais. 2015, f.103.
Dissertation (Postgraduate degree in Process Technology and Engineering) University of
Caxias do Sul, Caxias do Sul, 2015.
21. TREVISAN, V. ProposedMethodology for Determining Specific Activity Hidrognica. 2010,
187 f. Dissertation(Graduate in Water and Environmental Sanitation) University Federal of
Rio Grande do Sul, Porto Alegre, 2010.
22. ZHANG, C. et al.Reviewing the anaerobic digestion of food waste for biogs production.
Renewable and Sustainable Energy Reviews, 38, 383392, 2014.

10