MATTU UNIVERSITY

COLLAGE OF NATURAL SCIENCE AND COMPUTIONAL

DEPARTMENT OF BIOLOGY

BIOGAS PRODUCTION FROM MUSHROOM SUBSTRATE IN BOTO

KEBELE, MATTU TOWN

BY
1. SAHILUA BEHAYILU………………..……………………………RU2228/14
2. LIYA MARU………………………………………………………..RU1684/14

ADVISOR: LEMA EETEFA (MSC., ASSISTANT PROFESSOR)

A PROPOSAL SUBMITTED TO DEPARTMENT OF BIOLOGY IN PARTIAL

FULFILLMENT OF THE REQUIREMENT FOR THE COURSE RESEARCH
METHOD AND REPORT WRITING IN SCIENCE

NOVEMBER, 2024
METTU, ETHIOPIA

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 TABLE OF CONTENTS

1. Background of the study..............................................................................................................1

1.1. Statement of the problem..................................................................................................2

1.2. Significance of the study...................................................................................................2

1.3. Objectives..........................................................................................................................2

1.3.1. General objectives.....................................................................................................2

1.3.2. Specific objective..........................................................................................................2

2. LITERATURE REVIEW.........................................................................................................4

2.1 Basics of anaerobic digestion.................................................................................................4

2.2 Biochemical reactions in anaerobic digestion...................................................................4

2.3 Factors that affect anaerobic digestion..............................................................................5

2.4 Temperature......................................................................................................................5

2.5 pH......................................................................................................................................5

2.6 Inhibitory substances.........................................................................................................6

2.7 Anaerobic digestion technologies.....................................................................................6

2.8 Block scheme of anaerobic digestion and biogas/digestate utilisation.............................6

2.9 Biogas production, storage, treatment and use....................................................................7

2.10 The anaerobic digestion residue management..................................................................8

3. MATERIAL AND METHODS..............................................................................................12

3.1. Description the study area...................................................................................................12

3.2. Study Design.......................................................................................................................12

3.3. Methods of data collection..................................................................................................13

3.4. Methods of data analysis.....................................................................................................13

4.0 BUDGET BREAKDOWN AND WORK PLAN....................................................................14

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 4.1 Time Schedule......................................................................................................................14

4.2 Budget cost...........................................................................................................................15

5. REFERENCES..........................................................................................................................16

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 1. INTRODUCTION

1.1. Background of the study

Biogas is typically refers to a mixture of different gases produced by the breakdown of organic
matter in the absence of oxygen. It can be produced by row materials such as agricultural waste,
manure, municipal waste, plant material, sewage, green waste or food waste. It is commonly
used to refer a gas has been produced by the biological break down of organic matter in the
absence of oxygen. The organic material in the substrate is reduced and converted to Biogas by
microorganisms. It also takes place in an oxygen free environment Biogas Technology is the
application of the process that is based on the bacteria fermentation of organic materials, in the
absence of Air, to produce Flammable gas that can be put to various end uses. In practice the
organic material normally used include manure from animals (catle, pigs, goats, and poultry).
Household or market garbage, waste waters and waste of crop or agro industrial origin. These
materials are usually subjected to anaerobic (without air) fermentation in a biogas plant and the
gas produced is known as biogas.

Chemically biogas composed mainly of 60% methane or (flammable component) and 40%
carbon dioxide. The benefits of biogas technology at the community level include the utilization
of biogas for coking, water heating, andg, lighting's raining of kerosene refrigerators and
brooding of chicks or piglets. When produced in large quantities, biogas can also be used to
generate Electric city. Additional the fermented manure residues from the biogas plant contain
significant amount of nitrogen, phosphorus and potassium and can thus be used as organic
fertilizer for a variety of crops. Residues can also be used in fish production (agriculture) and
mushroom growing. Biogas technology has, there for, the potential to all viote poverty providing
substitutes for expensive fuels and commercial fertilizers, improving agri/aqua-cultural yields,
reducing local deforestation, treating jobs and income as well as strengthening the indigenous
technological knowhow.

The base line study conducted by the Department of energy in 1993. Fire wood and charcoal,
collectively known as wood fuel are the major source of energy supply in Zambia accounting are
about 80% of the total national energy consumption. Wood fuel has in most cases been realized

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by in discernment cutting of large tracts of forest areas which results in deforestation and then
environmental degradation. It is in the light of the above that build during and industrial research
unit (BIMU) of national institute for scientific and research (NISIR) in an effort to increase the
energy mix for the households in the nation started doing research in biogas technology.
According to 2008 study collected by the science and biogas children magazine. Methane biogas
from cow manure would be sufficient to produce 100 billion kilowatt hours enough to power
millions of homes across America.

1.1. Statement of the Problem

This study will be conducted on the production of biogas from mushroom substrate in Boto
kebele. Biogas production is a little beat long cross to get its products. The present study will be
under taken to utilize the waste generated during mush room sown production for biogas
production and the wastes generated during mush room cultivation process for the multiplication
of biogas trichoderma horzianum. The main problem of this study will be to disseminate the
result of established technologies on biogas production from mush room substrate with in
absence of oxygen.

1.2. Significance of the Study

The main importance of this study will be as follows:-
 To identify the mush room substrate they are mainly used for biogas production.
 To provide a brief description on the production of biogas from mush room substrate.
 To understand how biogas does is produce from mush room substrate with in absence of
oxygen.

1.3. Objectives
1.3.1. General Objectives
To established technologies for integration of mushroom cultivation (substrate) with biogas
production.

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1.3.2. Specific Objectives
 To identify mushrooms appropriate for biogas production
 To explore evaluate coffee wastes, teff, ficus vasta wastes and mixture of cow dung and
chickens waste.
 To evaluate the feasibility of developed technology for biogas production.

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 2. LITERATURE REVIEW
Biodegradable organic waste can be treated with or without air access. Aerobic process is
composting and anaerobic process is called digestion. Composting is a simple, fast, robust and
relatively cheap process producing compost and CO2 (Chiumenti et al. 2005, Diaz et al. 2007).
Digestion is more sophisticated, slow and relatively sensitive process, applicable for selected
input materials (Polprasert, 2007). In recent years anaerobic digestion has become a prevailing
choice for sustainable organic waste treatment all over the world. It is well suited for various wet
biodegradable organic wastes of high water content (over 80%), yielding methane rich biogas for
renewable energy production and use.

2.1. Basics of Anaerobic Digestion

This section deals with anaerobic waste treatment methods only, as the most advanced and
sustainable organic waste treatment method. Anaerobic digestion (WRAP 2010) is “a process of
controlled decomposition of biodegradable materials under managed conditions where free
oxygen is absent, at temperatures suitable for naturally occurring mesophilic or thermophilic
anaerobic and facultative bacteria and archaea species, that convert the inputs to biogas and
whole digestate“. It is widely used to treat separately collected biodegradable organic wastes and
waste water sludge, because it reduces volume and mass of the input material with biogas
(mostly a mixture of methane and CO2 with trace gases such as H2S, NH3 and H2) as by-product.
Thus, anaerobic digestion is a renewable energy source in an integrated waste management
system. Also, the nutrient-rich solids left after digestion can be used as a fertilizer.

2.2 Biochemical reactions in anaerobic digestion

In most cases biomass is made up of large organic compounds. In order for the microorganisms
in anaerobic digesters to access the chemical energy potential of the organic material, the organic
matter macromolecular chains must first be broken down into their smaller constituent parts.
These constituent parts or monomers such as sugars are readily available to microorganisms for
further processing. The process of breaking these chains and dissolving the smaller molecules
into solution is called hydrolysis. Therefore hydrolysis of high molecular weight molecules is the
necessary first step in anaerobic digestion. It may be enhanced by mechanical, thermal or

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chemical pretreatment of the waste. Hydrolysis step can be merely biological (using hydrolytic
microorganisms) or combined: bio-chemical (using extracellular enzymes), chemical (using
catalytic reactions) as well as physical (using thermal energy and pressure) in nature.

2.3 Factors that affect anaerobic digestion

As with all biological processes the optimum environmental conditions are essential for
successful operation of anaerobic digestion. The microbial metabolism processes depend on
many parameters; therefore these parameters must be considered and carefully controlled in
practice. Furthermore, the environmental requirements of acidogenic bacteria differ from
requirements of methanogenic archaea. Provided that all steps of the degradation process have to
take place in one single reactor (one-stage process) usually methanogenic archaea requirements
must be considered with priority. Namely, these organisms have much longer regeneration time,
much slower growth and are more sensitive to environmental conditions then other bacteria
present in the mixed culture.

However, there are some exceptions to the case:

2.4 Temperature
Anaerobic digestion can operate in a wide range of temperature, between 5°C and 65°C.
Generally there are three widely known and established temperature ranges of operation:
psychrophilic (15-20°C), mesophilic (30-40°C) and thermophilic (50-60°C). With increasing
temperature the reaction rate of anaerobic digestion strongly increases. For instance, with ideal
substrate thermophilic digestion can be approx. 4 times faster than mesophilic. However using
real waste substrates, there are other inhibitory factors that influence digestion, that make
thermophilic digestion only approx. 2 times faster than mesophilic. The important thing is, when
selecting the temperature range, it should be kept constant as much as possible. In thermophilic
range (50-60°C) fluctuations as low as ±2°C can result in 30% less biogas production (Zupančič
and Jemec 2010).
2.5 pH
In anaerobic digestion the pH is most affecting the methanogenic stage of the process. pH
optimum for the methanogenic microorganisms is between 6.5 and 7.5. If the pH decreases
below 6.5, more acids are produced and that leads to imminent process failure. In real digester

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systems with suspended biomass and substrate containing suspended solids, normal pH of
operation is between 7.3 and 7.5. When pH decreases to 6.9 already serious actions to stop
process failure must be taken. When using UASB flow through systems (or other systems with
granule like microorganisms), which utilize liquid substrates with low suspended solids
concentration normal pH of operation is 6.9 to 7.1. In such cases pH limit of successful operation
is 6.7. In normally operated digesters there are two buffering systems which ensure that pH
persists in the desirable range:

2.6 Inhibitory substances

In anaerobic digestion systems a characteristic phenomenon can be observed. Some substances
which are necessary for microbial growth in small concentrations inhibit the digestion at higher
concentrations. Similar effect can have high concentration of total volatile fatty acids (tVFA’s).
Although, they represent the very substrate that methanogenic archaea feed upon the
concentrations over10,000 mgL-1 may have an inhibitory effect on digestion (Mrafkova et al.,
2003; Ye et al., 2008). Inorganic salts can significantly affect anaerobic digestion. In real
operating systems it is unlikely that inhibitory concentrations of inorganic salts metals would
occur, mostly because in such high concentrations insoluble salts would precipitate in alkaline
conditions, especially if H2S is present. The most real threat in this case is sodium inhibition of
anaerobic digestion. This can occur in cases where substrates are wastes with extremely high salt
contents (some food wastes, tannery wastes…) or when excessive use of sodium substances were
used in neutralization of the substrate or the digester liquid. Study done by Feijoo et al. (1995)
shows that concentrations of 3000 mgL-1 may already cause sodium inhibition.
2.7 Anaerobic digestion technologies
Block scheme of anaerobic digestion that technological process of typical anaerobic digestion. It
consists of three basic phases:
i) substrate preparation and pre-treatment,
ii) anaerobic digestion and
iii) post treatment of digested material, including biogas use. In this section all of the
processes will be elaborated in detail.

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2.8 Block scheme of anaerobic digestion and biogas/digestate utilisation
Thermal pre-treatment rewards with up to 30 % more biogas production if properly applied. This
process occurs at temperature range of 135-220°C and pressures above 10 bar. Retention times
are short (up to several hours) and hygienisation is automatically included. Pathogenic
microorganisms are completely destroyed. The process runs economically only with heat
regeneration. When heat is regenerated from outflow to inflow of the pre-treatment process, it
takes only slightly more heat than conventional anaerobic digestion. Such process is very
appropriate for cellular material such as raw sewage sludge. It is also possible to use biological
processes as pretreatment. They are emerging in the world. Disintegration takes place by means
of lactic acid which decomposes complex components of certain substrates. Recently also
disintegration with enzymes has been quite successful, especially using cellulose, protease or
carbohydrases at a pH of 4.5 to 6.5 and a retention time of at least 12 days, preferably more
(Hendriks and Zeeman 2009).
Many research data have been published giving considerable attention to this kind of processes
(Dinsdale et al., 2000; Song et al., 2004; De Gioannis et al., 2008; Ponsá et al., 2008). Both
stages can be either mesophilic or thermophilic, however it is preferred that the hydrolysis-
acidogenic reactor is thermophilic and methanogenic is mesophilic. Typical HRT for the
thermophilic hydrolysis-acidogenic reactor is 1-4 days, depending on the substrate
biodegradability. Typical HRT for the methanogenic reactor is 10 - 15 days (mesophilic) and 10
- 12 days (thermophilic).
Advantages of this process beside shorter HRTs are higher organic load rate (20 % or more).
Many authors also report slightly better biogas yields (Messenger et al., 1993; Han et al., 1997;
Roberts et al., 1999; Tapana and Krishna, 2004). The only disadvantage is more sophisticated
equipment and process control, yielding the operation more expensive.

2.9 Biogas production, storage, treatment and use

When operating a biogas plant, biogas is the main product and considerable attention must be
given to its production, storage, treatment and use. Biogas production completely depends on the
efficiency of the anaerobic digestion and its microorganisms. Previous sections have shown what

9
conditions must be met to successfully operate anaerobic digestion. There are two distinct
parameters that describe the biogas production
1. Specific Biogas Productivity -SBP (it’s also called biogas yield). It is defined as volume of
biogas produced per mass of substrate inserted into digester (m3kg-1). There are variations; SBP
can be expressed in m3 of gas per kg of substrate: i) (wet) mass, ii) total solids, iii) volatile
organic solids or iv) COD. SBP tells us how much biogas was produced from the chosen unit of
substrate. Maximum possible SBP for certain substrate is called biogas potential. Biogas
potential can be determined by a standard method (ISO 1998).

2. Biogas Production Rate –BPR. It is defined as volume of biogas produced per volume of the
digester per day (m3m-3d-1). BPR tells us how much biogas we can gain from the active volume
of a digester in one day. SBP values of an optimally operating digester reach 80-90 % of the
biogas potential. Low pressure biogas holders arise in many variations. It is possible to include
biogas holder in the design of the digester. The most known is the digester with a movable cover.
These digesters are less common, because a movable cover requires increased investment and
operating expenditure. More common are external biogas holders that are widely commercially
available. Low pressure biogas holders require an extensive volume of 30 to 2000 m3 (Deublin
and Steinhauser, 2008).

After cleaning, biogas is used to produce energy. The most common way is to us all biogas in
cogeneration plant in CHP unit to produce power and heat simultaneously. In this case we can
achieve maximum power production and enough excess heat to run the digesters. The energy
required for heating the digester is also called parasitic energy. The anaerobic digesters require
heat to bring the substrate to operating temperature and to compensate the digester heat losses.
The digester also requires energy for mixing, substrate pumping and pre-treatment. The largest
portion of heating demands in the digester operation is substrate heating. It requires over 90 % of
all heating demands, and only up to 10 % is required for heat loss compensation (Zupancic and
Ros 2003).

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Consequently the whole biogas production process can be more economic, in some cases even
without considerable subsidies as well as more renewable energy is put to the energy supply.
Also in most cases, the investment costs of biogas plants may be less, since there is no CHP
plant. In order to be able to inject the biogas into natural gas as biomethane (Ryckebosch et al.,
2011) grid certain purity standards must be fulfilled, which in EU are determined by national
ordinances (a good example is the German ordinance for Biogas injection to natural gas grids
from 2008), where responsibilities of grid operators and biogas producers are determined as well
as quality standards are prescribed (DVGW, 2010). When injecting biomethane into the natural
gas grid some biogas must be used for the reactors self-heating.

2.10 The anaerobic digestion residue management

A quality management system (QMS) specific to a defined digestion process and its resulting
whole digestate or any separated liquor and separated fibre, should be established and
maintained. Anaerobically digested slurry or sludge contains 2-12 % of solids; wet waste from
solid state digestion contains 20-25 % solids. The digestate contains not degraded organic waste,
microorganism cells and structures formed during digestion, as well as some inorganic matter.
This is potentially an alternative source of humid material, nutrients and minerals to the
agricultural soil (PAS, 2010). It may be used directly or separated into liquid and solid part. The
liquid digestate is often recycled to the digestion process; some pre-treatment may be required to
reduce nitrogen or salt content. Freshly digested organic waste is not stable under environmental
conditions: it has an unpleasant odour, contains various noxious or corrosive gases such as NH3
and H2S, and still retains some biodegradability. In certain periods of a year it may be used in
agriculture directly, in most cases however it must be stabilized before being applied to the
fields.

Aerobic treatment (composting) is an obvious and straightforward solution to this problem. The
composting procedure has several positive effects: stabilization of organic matter, elimination of
unpleasant odours and reduction of pathogenic microorganisms to an acceptable level.
Composting, applied prior to land application of the digested waste, contributes also to a
beneficial effect of compost nitrogen availability in soil. (Zbytniewski and Buszewski, 2005;

11
Tarrasón et al., 2008). The simplest way is composting of the dehydrated fresh digestate in a
static or temporarily turned-over pile. A structural material is necessary to provide sufficient
porosity and adequate air permeability of the material in the pile. Various wood or plant
processing residues may be used as a structural material like woodchips, sawdust, tree bark,
straw and corn stalks provided that the sludge : bulk agent volume ratio is between 1:1 and 1:4
(Banegas et al., 2007). The final compost quality depends on the content of pollutants such as
heavy metals, pathogenic bacteria, nutrients, inert matter, stability etc. in the mature compost.
Typical quality parameters are presented in Table 1. The properties of the compost standard
leachate may also be considered. Heavy metals and persistent organic pollutants accumulate in
the compost and may cause problems during utilization. Compost quality depends on quality of
the input material, which should be carefully controlled by input analysis. Pathogenic bacteria
may originate from the mesophilic digestates or from infected co-composting materials, if
applied (e.g. food waste). If thermophilic phase period of the composting process has lasted at
least few days, the compost produced may be considered sanitized and free of pathogens such as
Salmonella, Streptococci and coliforms. The third important factor is presence of nitrogen.
Several authors have reported that the optimal C/N ratio is between 25/1 and 30/1 although
operation at low C/N ratios of 10/1 are also possible. With such low C/N ratios the undesirable
emission of ammonia can be significant (Matsumura et al., 2010). Characteristic values of
organic matter content and total nitrogen in the digested sludge are 50-70% and 1.5-2.5%,
respectively. In the first week of the digested sludge composting the total carbon is reduced by
between 11% and 27% and total nitrogen is reduced by between 13% and 23% (Pakou et al.,
2009; Yañez et al., 2009).

Highest degradation rates in the compost pile are achieved with air oxygen concentration above
15% which also prevents formation of anaerobic zones. The quality of aeration depends
primarily on structure and degree of granulation of the composting material; finer materials
generally provide better aeration of the compost pile (Sundberg and Jönsson, 2008). In the first
stages of degradation, acids are generated, and these tend to decrease the pH in the compost pile.
The optimum pH range for microorganisms to function is between 5.5 and 8.5. Elevated
temperature in the compost material during operation is a consequence of exothermic organic

12
matter degradation process. The optimum temperature for composting operation, in which
pathogenic microorganisms are sanitised, is 55-70°C. In the initial phases of composting the
prevailing microorganisms are fungi and mesophilic bacteria, which contribute to the
temperature increase and are mostly sanitised in the relevant thermophilic range. When
temperature falls many of the initial mesophilic microorganisms reappear, but the predominant
population are more highly evolved organisms such as protozoa and arthropods (Schuchard,
2005). For optimum composting operation the correct conditions must be established and are
determined by particle size distribution and compost pile aeration have shown that the air gaps in
the compost pile can be reduced from an initial 76.3% to a final 40.0%. The optimum moisture
content in the compost material is in the range of 50-70%. In the recent years the composting
practice for anaerobic digestate has been thoroughly studied for many different types of
substrates, for co-composting and with many different bulk agents (Nakasaki et al., 2009;
Himanen et al., 2011).

From various reasons the composting of the digestate residue is sometimes not possible (lack of
space, problems with compost disposal etc.). Alternatively the digestate may be treated by
thermal methods, which require higher solid content. Mechanical dehydration by means of
continuous centrifuges provides solid content about 30 % with positive calorific value.
Incineration may be carried out in a special kiln (most often of fluidized bed type) or together
with municipal waste in a grit furnace. Co-incineration in industrial kilns usually require drying
of sludge to 90 % dryness that gives calorific value of about 10 MJ/kg. Thermal methods are
more expensive than composting due to high energy demand for dehydration and drying,
sophisticated processes involved and strict monitoring requirements. Good review of the modern
alternative processes of anaerobic sludge treatment is presented by Rulkens (2008).

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 3. MATERIAL AND METHODS
3.1. Description the study area
The study will be conducted in Botu kebele Mettu town. Mettu town is found in west Illuababor
zone of Ethiopia. It is located about the 605 km south west from Addis Ababa and 5km from
west of Mettu.

3.2. Study Design

The study will be conducted on the production of biogas from mushroom substrate. The
experiment will be conduct to study utilization of wastes such as coffee waste, teff, ficus vasta,
gravel and a mixture of dung and chicken wastes. During the experimental work, 200g, 150g and
50g of coffee and cow dung, were respectively mixed with 100ml of water at the same time and
also 150g of teff and 50g of chickens were also mixed with 100ml of water. In addition of this a
mixture of 150gm ficus and 50gm cow dung were mixed in 100ml of water then again a mixture
of 150gm of ficus and 50gm chicken waste were mixed the same rate of 100ml water. Lastly
200gm, 150gm and 50gm of gravel and cow dung were respectively mixed with 100ml of water
and at the same times a mixture of 150g gravel and 50g from chicken were mixed with 100ml of
water
3.3. Methods of data collection
Material used for data collection are mainly note book, pen,and recorder/smart mobile. In this
study design, the data will be collected by using primary data collection techniques or also that

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data will be collected by mycology or microbiology research lab, or data collected from Botu
kebele. It is also collected from the area by using primary data collection and its surroundings.

3.4. Methods of data analysis

The collected data will be analyzed or infergarate by using descriptive statistics. After the all
necessary data will method like procedure, table and figure distribution would be used during the
experiment.

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 4. BUDGET BREAKDOWN AND WORK PLAN

4.1 Time Schedule

Time Schedule is a time expected to accomplish research proposal and time table that the
researcher will expect to carry out activity in terms of month and expected to complete the study.
Table 1: Time schedule of the research

N activities octob novemb decemb janua februa marc apr ma jun

o er er er ry ry h il y e
1 Select a 
title and
approval
2 Identify 
the
problem
s
3 Data 
collectio
n
4 data 
analysis
5 Proposal 
submissi
on
6 Proposal 
defense
7 Writing 
thesis
8 Submissi 
on of
final
paper
9 Work on 
commen

16
 t
1 Defense 
0 thesis

4.2 Budget cost

To accomplish this activity, it requires budget that will be paid for personal, materials and
contingences as follows.
Table 2: Budget breakdown
No Item Quantity Unit Unit in price Total price
1 CD-RW 3 Paper 30 90
2 Photo copy 25-35 Pages 1 35
3 Transport - Km 12 30
4 Secretary 2 No 100 100
5 Paper 1 Ream 150 150
6 Pen 5 Pieces 10 50
7 Key 6 No 30 180
informants
8 Total 635

To conduct this research totally 635 birr is important.

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5. REFERENCES
1. Banegas V., J.L. Moreno, J.I. Moreno, C. Garcia, G. Leon, T. Hernandez,
(2007).Composting.
2. Anaerobic and aerobic sewage sludges using two proportions of sawdust. Waste Manag. 27,
1317-27.
3. De Gioannis G., Diaz L.F., Muntoni A., Pisanu A., (2008). "Two-phase anaerobic digestion
within a solid waste/wastewater integrated management system." Waste Management
28(10): 1801-1808.
4. Deublein, D. and A. Steinhauser (2008). Biogas from waste and renewable resources.
Weinheim, Willey-VCH Verlag GmbH & Co. KGaA.
5. Dinsdale R. M., Premier G.C., Hawkes F.R., Hawkes D.L., (2000). "Two-stage anaerobic
co-digestion of waste activated sludge and fruit/vegetable waste using inclined tubular
digesters." Bioresource Technology 72(2): 159-168.
6. DVGW - Deutscher Verein des Gas- und Wasserfaches, DVGW G 262 “Nutzung von Gasen
aus regenerativen Quellen in der der Öffentlichen Gasversorgung” (2010), available online
with limited access at http://www.dvgw.de/gas/gesetze-und-erordnungen/.
7. Feijoo G., Soto M., Méndez R., Lema J.M., (1995). "Sodium inhibition in the anaerobic
digestion process: antagonism and adaptation phenomena." Enzym Microb Technol 17(2):
180-188.
8. GNS (2009) Nitrogen removal from manure and organic residues by ANAStrip – process
(System GNS). http://www.gns-halle.de/english/site_1_6.htm (Access 11th August 2011).
9. Han Y., Sung S., Dague R.R., (1997). "Temperature-phased anaerobic digestion of
wastewater sludges." Water Sci Technol 36(6-7): 367-374.
10. Hendriks A.T.W.M., Zeeman G., (2009). "Pretreatments to enhance the digestibility of
lignocellulosic biomass." Bioresour Technol 100(1): 10-18.
11. Himanen M., Hänninen K., (2011). Composting of bio-waste, aerobic and anaerobic
sludges– Effect of feedstock on the process and quality of compost. Bioresource
Technology, Volume 102, Issue 3, p. 2842-2852.

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12. ISO (1998) EN ISO 11734 (1998) Water Quality – Evaluation of the »Ultimate« Anaerobic
Biodegradability of Organic Compounds in Digested Sludge – Method by Measurement of
the Biogas Production, International Standard Organization.
13. Messenger J., de Villers H.A., Laubscher S.J.A., Kenmuir K. and Ekama G.A. (1993).
"Evaluation of the Dual Digestion System: Part 1: Overview of the Milnerton Experience."
Water SA 19(3): 185-192.
14. Mrafkova L., Goi D., Gallo V., Colussi I., (2003). "Preliminary Evaluation of Inhibitory
Effects of Some Substances on Aerobic and Anaerobic Treatment Plant Biomasses." Chem
Biochem Eng Q 17(3): 243-247.
15. Nakasaki K., Tran L.T.H., Idemoto Y., Abe M., Rollon A.P., (2009). Comparison of organic
matter degradation and microbial community during thermophilic composting of two
different types of anaerobic sludge. Bioresource Technology, Volume 100, Issue 2, p. 676-
682.
16. Polprasert C. (2007) Organic Waste Recycling – Technology and Management, 3rd Ed., e-
book, IWA Publishing , London Ponsá S., Ferrer I., Vázquez F., Font X. (2008).
"Optimization of the hydrolytic-acidogenic anaerobic digestion stage (55 °C) of sewage
sludge: Influence of pH and solid content." Water Res 42(14): 3972-3980.
17. Roberts R., Davies W.J., Forster C.F., (1999). "Two-Stage, Thermophilic-Mesophilic
Anaerobic Digestion of Sewage Sludge." Process Saf Environ Protec 77(2): 93-97.
18. Rulkens W., (2008). Sewage Sludge as a Biomass Resource for the Production of Energy:
Overview and Assessment of the Various Options, Energy Fuels, 22 (1), pp 9–15
19. Ryckebosch E., Drouillon M., Vervaeren H., (2011). “Techniques for transformation of
biogas to biomethane”, Biomass and Bioenergy, 35(5), p. 1633-1645.
20. Schuchard F., (2005). Composting of organic waste. In: Enviromental biotechnology
concepts and aplications. Editors:Jördering, H.J., Winter, J. Weinheim: Willey-VCH Verlag,
p. 333-354.
21. Sežun M., Grilc V., Zupančič G.D. and Marinšek-Logar R., (2011). Anaerobic digestion of
brewery spent grain in a semi-continuous bioreactor: inhibition by phenolic degradation
products. Acta chim. slov.., 58, 1, 158-166.

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22. Song Y.C., Kwon S.J., Woo J.H., (2004). "Mesophilic and thermophilic temperature co-
phase anaerobic digestion compared with single-stage mesophilic- and thermophilic
digestion of sewage sludge." Water Research 38(7): 1653-1662.
23. Sundberg C., Jönsson H., (2008). Higher pH and faster decomposition in biowaste
compostingby increased aeration. Waste Management, Volume 28, Issue 3, p. 518-526.
24. Tapana C., Krishna P.R., (2004). "Anaerobic Thermophilic/Mesophilic Dual-Stage Sludge
Treatment." Journal of Environmental Engineering 126(9): 796-801.
25. WRAP (2010) Specification for whole digestate, separated digestate, separated liquor and
separated fibre derived from the anaerobic digestion of source-segregated biodegradable
materials, Publicly Available Specification (PAS) 110, U.K.
26. Yañez R., Alonso J.L., Díaz M.J., (2009). Influence of bulking agent on sewage sludge
composting process, Bioresource Technology, Volume 100, Issue 23, p. 5827-5833.
27. Ye C., Cheng J.J., Creamer K.S., (2008). "Inhibition of anaerobic digestion process: A
review." Bioresource Technology 99(10): 4044-4064.
28. Zupančič G. D., Roš M., (2003). "Heat and energy requirements in thermophilic anaerobic
sludge digestion." Renewable Energy 28(14): 2255-2267.
29. Zbytniewski, R., Buszewski, B., 2005. Characterization of natural organic matter (NOM)
derived from sewage sludge compost. Part 1: chemical and spectroscopic properties.
Bioresource Technology, Volume 96, Issue 4, p. 471-478.

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