This thesis comprises 30 ECTS credits and is a compulsory part in the Master of Science with a Major in Environmental Engineering, 120 ECTS credits No. 8/2009 Application of Various Pretreatment Methods to Enhance Biogas Potential of Waste Chicken Feathers Azar Khorshidi Kashani
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Application of Various Pretreatment Methods to Enhance Biogas Potential of Waste Chicken Feathers
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This thesis comprises 30 ECTS credits and is a compulsory part in the Master of Science with a Major in Environmental Engineering, 120 ECTS credits
No. 8/2009
Application of Various Pretreatment Methods to Enhance Biogas Potential
of Waste Chicken Feathers
Azar Khorshidi Kashani
Application of various pretreatment methods to Enhance Biogas Potential of Waste Chicken feathers
action are utilizing in many energy fields such as electricity generation, transportation
fuels, industrial processes, heating , cooling and process steam. Although renewables
currently provide less than 10% of the world's energy, renewable energy sources have the
potential to exceed current global energy demands even with existing technologies [1].
13
1.1.2 Biomass
Biomass as a major source of renewable energy accounts for about 14% of primary
energy consumption, and following oil, coal and natural gas is the fourth world-wide
energy resource. The world production of biomass is estimated at 146 billion metric tons
a year, mostly coming from wild plant growth [4,5].
The major resources of biomass are agricultural crops, plants and forestry residues,
organic components of municipal and industrial wastes and even the fumes from
landfills. Biomass can be converted to non-solid fuels form including liquid biofuel
(bioethanol and biodiesel) and gaseous biofuels (biogas, syngas,…). Fig. 3 Indicated
potential pathway for biofuel production.
Fig. 3. Potential pathway for biofuel production [6].
1.2 Biogas Biogas is the gaseous biofuel made through anaerobic digestion process or fermentation
of organic fraction of biomaterials. Biogas can be also captured from landfills. Almost all
kinds of organic and biodegradable materials such as municipal and industrial organic
wastes, sludge from sewage treatment plants and process water from the food industry,
energy crops and crop residues can be utilized as the resources for biogas production.
Biogas comprises from methane (CH4), carbon dioxide (CO2) and trace amounts of some
other components. Table 1 shows the typical composition of biogas.
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compound Percentage (%)
Methane, CH4 50-75
Carbon dioxide, CO2 25-50
Nitrogen, N2 0-10
Hydrogen, H2 0-1
Hydrogen sulfide, H2S 0-3
Oxygen, O2 0-2
Table 1.Typical composition of biogas [7]. 1.2.1 Biogas applications and benefits
Biogas is an environmentally friendly, clean, cheap and versatile fuel. Anaerobic
digestion substrate for biogas production can be obtained from almost all kinds of bio-
wastes and non-food based biomasses. Therefore biogas has no potential negative impact
on food chain products and prices, changes in land use and deforestation [8,9]. Combustion of biogas has less dangerous and neutral carbon dioxide emissions [10].
Moreover methane is a potent greenhouse gas, and hence capturing and burning it helps
environment from the global warming point of view. Biogas has a wide range of
applications e.g. in transportation, electricity production, cooking, space heating, water
heating and industrial process heating or even as a renewable feedstock to produce
hydrogen [8]. Table 2 shows some typical applications for one cubic meter of biogas.
Application 1m3 biogas equivalent
Lighting Cooking Fuel replacement Shaft power Electricity generation
equal to 60 -100 watt bulb for 6 hours can cook 3 meals for a family of 5 - 6 0.7 kg of petrol can run a one horse power motor for 2 hours can generate 1.25 kilowatt hours of electricity
Table 2. Some biogas equivalents [11,12].
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Europe seems to be the leader in the global production and use of biogas [10]. Fig. 4 shows
the deployment of anaerobic digestion in the EU and the world from 1995 to 2010.
Fig. 4. Deployment of anaerobic digestion in the EU and the world [9].
UK studies have shown that biogas is much cleaner and more efficient than biofuels for
use in transport. According to an EU well-to-wheel study of more than 70 different (fossil
and renewable) fuels and energy paths, biogas is the cleanest and most climate-neutral
transport fuel of all [10]. “A natural gas vehicle reduces CO2 over a gasoline car by 20-
30%. A car running on bio-methane reduces CO2 on a well-to-wheel basis by more than
100%over a petroleum-fuelled car [8].”
Biogas along with fossil natural gas is currently fuelling over 800,000 cars, truck and
buses in Europe and nearly 8 million vehicles worldwide [8]. Compressed biogas
is becoming widely used in vehicles in Sweden, Switzerland and Germany [7]. “Sweden
has led the world in the usage of biogas in transportation since 1996. Biogas producers
are operating a fleet of city buses in Sweden. Strong government support is important, it
includes 30 percent investment support, zero tax, reduced income tax for company car
users, and no congestion fees in the capital city of Stockholm [1].”
Among biomass sub-sectors, solid biomas (72.5% biomass electricity) has increased by
an avarage of 5.8% per year from 1997 to 2007. However, growth in biogas electricity
has been much more considerable (an average of + 12.9% per year) [14]. European
Biogas electricity production in 2006 was 17272GWh per year, of which 7338GWh was
produced by Germany alone [15]. Beside biogas, anaerobic digestion produces high
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nutrient content fertilizers to use in agriculture [11]. Furthermore, biogas production has
no geographical limitations and doesn’t need sophisticated technology [16]. Biogas can
be produced even by a very basic construction using mostly used materials providing a
few simple design rules are followed. Moreover, biogas production is possible in small
scale sites, to obtain for outlying areas [17]. Accordingly, biogas is a 100% sustainable
fuel playing also a very important role in environmental friendly waste management and
organic waste disposal [8].
1.2.2 Anaerobic Digestion Process
Anaerobic digestion process for generation biogas occurs in four steps: Hydrolysis,
Acidogenesis, Acetogenesis and Metanogenesis. In the first step, hydrolysis, insoluble
and complex organic compounds such as lipids, polysaccharides, proteins, fats, nucleic
acids, etc. transform into soluble and simpler organic materials such as amino acids,
sugars and fatty acids by strict anaerobic hydrolytic bacteria [18,19]. In the acidogenesis
step obligate and facultative anaerobic group of bacteria (acidogens) ferments and
breakdown soluble products from the first step into acetic acid, hydrogen, carbon dioxide,
some volatile fatty acids (VFA) and alcohols. In the third step, acetogenesis, long chain
fatty acids and volatile fatty acids will be converted to acetate, hydrogen and carbon
dioxide by obligate hydrogen-producing acetogens [18]. Finally in the methanogenesis
Various concentrations of Lime (Ca (OH)2 g/g TS F) were added to the mixtures of 2
different concentrations (40 &100g TS/l water) of milled and 105°C dried chicken
feathers. 50 ml of each sample was prepared in duplicate. Afterward, samples were
closed with aluminum foil loosely and were heated in the autoclave at different
temperatures for different treatment times according to the Table 5:
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Exp. Number
Feather Concentration (g TS F/l liquid)
Lime loading (g/g TS F)
Autoclave Temperature
(°C)
Time (min)
0.1 0.2
0.4 1 2
1
40
4
100, 110, 120
30,60,120
0.1 0.2 0.4 1
2
100
2
100, 110, 120
60,120
Table 5. Thermo-chemical treated samples and treatment conditions (Exps.1 and 2) After cooling the samples to the room temperature in a desiccator, pH measurement for
the samples was carried out. In general, due to the presence of the lime pH values of the
treated samples has been maintained around 11.5-12.5.
To adjust the pH of the samples to the suitable value for anaerobic digestion and also to
convert the existing lime in the samples to the water-soluble Ca(HCO3)2 (as much as
possible), samples were carbonated with pure CO2 gas while the pH were controlled
continuously. In this way the pH of the samples decreased to about 8-8.5 and major
amount of the lime was converted to water-soluble calcium bicarbonate (Ca(HCO3)2) and
also low soluble calcium carbonate (CaCO3) [35].
One of each duplicated samples were centrifuged and the liquid phase of them were used
for soluble chemical oxygen demand (SCOD) concentration measurement.
Considering the SCOD measurement results, the following uncentrifuged samples which
their centrifuged couples had revealed high SCOD concentration and also contented
much lower amount of the precipitated lime and calcium carbonate (CaCO3) were
selected to use for the anaerobic digestion process (samples had been made in 50ml
volume):
- For experiment 1, using 40 g TS feather/l concentration, the selected samples had been
treated under the following conditions:
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1- 0.1g lime /g TS feather, 30 min, 100°C:
2g TS feather + 48 ml water + 0.2g lime
2- 0.1g lime /g TS feather, 30 min, 120°C:
2g TS feather + 48 ml water + 0.2g
3- 0.2g lime /g TS feather, 1 h, 120°C:
2g TS feather + 48 ml water + 0.4g lime
4- 0.2g lime /g TS feather, 2 h, 120°C:
2g TS feather + 48 ml water + 0.4g lime
- For experiment 2, using 100 g TS feather/l concentration, the selected samples had been
treated under the following conditions:
1- 0.1g lime /g TS feather, 2h, 120°C:
5g TS feather + 45 ml water + 0.5g lime
2- 0.2g lime /g TS feather, 2h, 120°C:
5g TS feather + 45 ml water + 1g lime
3- 1g lime /g TS feather, 2h, 120°C:
5g TS feather + 45 ml water + 5g lime
4- 2g lime /g TS feather, 2h, 120°C:
5g TS feather + 45 ml water + 10g lime
3.4.2 Biological Pretreatments (Experiment 3)
In this series of experiment the effect of thermal, enzymatic, combined thermo-enzymatic
and combined chemo-enzymatic pretreatments on hydrolysis of feather were examined.
Milled and oven dried feathers, 0.9g TS F/vial, (1g F/vial) were pre-treated in the small
flasks (118 ml), in triplicate and one excess sample for SCOD measurement. For the
enzymatic treatment an alkaline endopeptidase enzyme, Savinase, was used.
Furthermore, for chemo-enzymatic treatment sodium sulfite was also added as chemical
reductant agent to cleavage disulphide bonds. The pH of the samples was adjusted to
pH=8.0 using phosphate buffer. The total volume of each sample was 10 ml.
Pretreatments were conducted using the following conditions and materials:
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1- Thermal treatment: autoclaving for 5min at 120°C
0.9g TS feather + 9.1 g potassium phosphate buffer solution
Fig. 20. Average maximum methane production curves for triplicate lime treated
sample of Exp. 2, during 15 days of incubation.
The increased SCOD concentration and meanwhile decreased methane yield of these
samples probably reflect the effect of the overloading of the system with organic
substrate leading in accumulation of ammonia which inhibited CH4 productivity of the
protein material during anaerobic digestion process. Accordingly, less feathers loading
may result in more efficient anaerobic digestion process.
It is mentionable that the same considerations i.e. high SCOD content, least precipitation
of lime and calcium carbonate (CaCO3) and optimal conditions had been applied in
selection of the samples of experiment 2 for anaerobic digestion process.
After lime treatment the measured pH for the samples was around 11.5-12.5. Carbonating
samples with CO2 gas before digestion process decreased the pH to about 8-8.5. Although
no buffer was used to adjust the pH during the digestion process, final measurement of
the pH indicated that the pH of the samples had been maintained almost at the same level
of the starting of the AD process (pH of 8-8.5). According to the literatures “Calcium
hydroxide is an alkaline material poorly soluble in water that maintains a relatively
constant pH (~12), provided enough lime is in suspension. This low solubility ensures a
constant pH during the thermo-chemical treatment and relatively weaker conditions
(compared to sodium hydroxide and other strong bases) that helps in reducing the
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degradation of susceptible amino acids. The new carboxylic acid ends react in the
alkaline medium to generate carboxylate ions, consuming lime in the process [35].”
Lo´pez Torres et al.,2007 [104] found also the similar point and reported that in contrast
with the other AD systems the digesters fed with lime pretreated waste maintained its
alkalinity and neutral pH during digestion process without necessity of continuously
addition of alkali.
Samples 2 and 3 of experiment 1 had a fast onset in methane production but samples 4
and 5 had a one week lag phase. However, in repetition of the AD process using liquid
phase of samples 4 and 5 no delay was observed in the start of the methane production.
The ammonia production of in vitro rumen digested lime soluble chicken feather keratin
was also previously studied by Coward-Kelly et al. (2005) [35]. They found that
ammonia production from soluble keratin in rumen fluid was similar to that of soybean
and cottonseed meals and was greatly less than that of urea. Soybean and cottonseed
meals are the most popular protein sources for cattle which do not result in ammonia
toxicity. Therefore, soluble feather keratin is likely more readily digested than the other
proteins and no ammonia toxicity will result from cattle being fed soluble keratin [35].
Similar performance might be expected from lime treated samples during anaerobic
digestion of feather for biogas production, namely no ammonia toxicity is produced and
inhibits the anaerobic microorganisms for the recommended condition.
According to Lo´pez Torres et al. 2007, Alkaline pretreatment of organic materials with
Ca(OH)2 not only increases the level of soluble COD but also surface area of complex
organic matter, due to fiber swelling. These facts make these materials more susceptible
to enzymatic attack by microorganisms and enhance anaerobic digestion processes [104].
Another significant advantage of alkaline treatment is disruption of the disulphide bonds
in feather which was previously noticed by Salminen et al. [30]. All of the above results support the positive effect of lime pretreatment on hydrolysis of
the chicken feather and other organic materials, and according to the achieved results in
the present study pretreatment of chicken feather under (40g TS feather/l, 0.1g Ca(OH)2/g
dry feather, 100°C, 30 min) condition is the optimum condition to exert the most
significant effect on increasing the methane yield of chicken feather. Coward-Kelly et
al., 2005 [35] found that pretreatment of feather under 0.1g Ca(OH)2/g dry F, 100°C and
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300 min treatment conditions can solublise 80% of feather keratin to produce an amino
acid rich foodstuff for animals and in this study the pretreatment times was modified to
30 min for anaerobic digestion of feathers resulted in 96.8 % of the theoretical potential
methane productivity. This shorter treatment time is safer for AD process and more
profitable from economical point of view.
5.3 Effect of biological treatments on SCOD concentration (Exp.3)
In this series of the experiments the effect of the thermal, enzymatic, combined thermal-
enzymatic and combined chemical-enzymatic pretreatment on solublisation and methane
yield of chicken feather were investigated.
Fig. 21 shows the samples after enzymatic, chemo-enzymatic and thermo enzymatic
pretreatment.
Fig. 21. Enzymatic, chemo-enzymatic and thermo-enzymatic pretreated samples (Exp.3).
The average values for SCOD concentration of the pretreated samples are demonstrated
in the Fig. 22.
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0
5000
10000
15000
20000
25000
30000
35000
40000
SCOD (mg/l)
Enzymatic Thermo-Enzymatic
Chemo-Enzymatic
Treated Samples
COD Concentration
Enzymatic
Thermo-Enzymatic
Chemo-Enzymatic
Fig. 22. Results of SCOD measurement for enzymatic and combined enzymatic
pretreated samples of Exp.3.
As seen in the Fig. 22 these methods of pretreatment solublised the feather and showed
positive effect on increasing the SCOD concentration of the samples. As seen in the
Fig.22 these methods of pretreatment solublised the feather and showed positive effect on
increasing the SCOD concentration of the samples. But in contrast to lime treatment,
where the highest relative SCOD release was around 1680 mg SCOD/g TS F here the
highest relative SCOD release value was 407 mg SCOD/g TS F produced by the chemo-
enzymatic treatments. It is still much lower than the relative SCOD release of 1040 mg
SCOD/g TS F for the recommended lime treatment conditions of 40g TS F/l liquid, 0.1g
Ca(OH)2/g TS F, 100°C and 30 min.
5.4 Effect of biological treatments on anaerobic digestion performance
Although combined enzymatic pretreatments could solublise feather and increase the
SCOD concentration, methane yield enhancement by these methods were also much
lower than those of lime pretreatment. Table 9 and Fig. 23 illustrate maximum methane
productivity of these samples during 50 days of anaerobic incubation:
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Table 9. Results of SCOD and average maximum methane yield of triplicate thermal, enzymatic and combined enzymatic pretreated samples of Exp.3.
Average Normal vol CH4 EXP6- Feather
-0.1
-0.05
0
0.05
0.1
0.15
0.2
0 10 20 30 40 50 60
Time (days)
N V
OL
(m3/
kg V
S) Untreated
Thermal
Enzymatic
Thermo-Enzymatic
Chemo-Enzymatic
Fig. 23. Average maximum methane production curves fort triplicate thermal, enzymatic
and combined enzymatic treated samples of Exp.3, during 50 days incubation.
samples
Feathers concentration
Treatment Conditions
SCOD Mg/L
Maximum Methane yield Nml/gVS
Percent of theoretical methane potential
1 Control, Untreated ---- 135 27.2%
2 Thermal, 120°C, 5 min
---- 143 28.8%
3
Enzymatic, 1%w enzyme/vial, 55°C, 2 h
18,640
154
31%
4
Thermal-Enzymatic, 120°C, 5min- 1%w enzyme/vial, 55°C, 24 h
methane potential and produced an average maximum of 41 Nml CH4/g VS i.e. 3 times
less than untreated sample and only 8% of the theoretical methane potential during 50
days of anaerobic incubation, likely due to the high degradation of some amino acids
under the effect of the pretreatment conditions which leaded to more and quick formation
of some inhibitory compounds (e.g. ammonia and H2S) during anaerobic digestion
process [41-43]. Further experiments must be performed to determine the inhibitory
agents and reasons for the low methane production of these samples, as well. Also the
effect of the treatment conditions such as temperature, reaction time, enzyme and sodium
sulfite loading, etc. on the anaerobic digestion performance of these samples should be
investigated in the future works.
As a whole, the results of the experiments performed in this study revealed that the less
feather loading results in more efficient anaerobic digestion process.
Therefore, considering the results of this study, simplicity of the treatment method and
also the low price of lime, lime treatment under the above mentioned optimal condition
can be suggested as the most feasible and the highest efficient pretreatment method to
enhance chicken feather methane potential through anaerobic digestion process.
5.6 Future work
In the present study anaerobic digestion for lime treated samples were carried out in a
batch mode. The effects of the lime treatment on the methane efficiency of the chicken
feathers can also be evaluated in a fill-and-draw or semicontinuous anaerobic digestion
process suggested in previous studies and literatures as a more efficient process than
batch system [105,103,106]. Application of this method in anaerobic digestion of lime
treated feather would also demonstrate the long time anaerobic digestion performance of
the treated samples.
Inhibitory agents for anaerobic digestion of thermal and enzymatic and combined
thermo-enzymatic pretreated samples and other probable reasons of the declining of their
methane productivity after 33 days, and also fast deviation in the methane yield of the
chemical enzymatic pretreated sample after 12 days should be determined through
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performing further experiments. Moreover, optimization of the feather loading can be
performed and then the effect of the variation of the other treatment condition such as
temperature, reaction time, enzyme and sodium sulfite loading, etc. on the anaerobic
digestion performance can be further investigated.
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APPENDICES APPENDIX A: Tables and Data Figures for the Results of TS% & VS%
Measurement:
Table 10. The recorded weighs during TS measurement and the results for the TS% of the samples.
Table 11. The recorded weighs during VS measurement and the results for the VS%
of the samples.
Sample
Weight of oven dried (105°C) crucibles (g)
Total weight of air dried sample and dried(105°C) crucible (g)
Total weight of oven dried sample and crucible (105°C) (g)
Weight of dried sample (105°C) (g)
TS%
1
48.78
51.25
51.04
2.2593
91.40
2
47.67
50.21
49.98
2.3120
91.12
3
48.90
51.37
51.15
2.2535
91.35
Sample
Weight of oven dried (550°C) crucibles (g)
Total weight of oven dried (105°C) sample and dried (550°C) crucible (g)
Total weight of burned sample and crucible (550°C) (g)
Weight of burned sample (g)
VS% of TS%
VS%
1
44.84
45.62
44.67
0.0066
99.15
90.62
2
44.67
45.41
44.85
0.0041
99.44
91.06
3
45.00
45.74
45.00
0.0041
99.44
91.29
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APPENDIX B:
B.1 Data Figures and Tables for the Results of GC Measurements for
Lime Treated Samples: Tables below shows the average volume of methane produced by lime treated samples
containing 40g TS feather/l liquid during 50 days incubation, under thermophilic