What is a Biogas

Biogas is a by-product produced from the fermentation of organic substances by the action of bacteria

Biogas is composed mainly of CH4(Methane) and CO2(Carbon Dioxide) with traces of H2S(Hydrogen Sulfide),
H2O(Water vapor) and H2(Hydrogen).

Literature

The decomposition of waste material during anaerobic digestion is caused by bacterial action
rather than high temperatures. It takes place in almost any biological environment, but is
favored by warm, wet and low oxygen conditions.
Anaerobic digestion also occurs in two major situations created by human activities:
__ Sewage (human waste) or animal manure.
__ Landfill gas produced by domestic refuse buried at landfill sites.

REQUIREMENTS FOR A SUCCESSFUL SYSTEM

1. Acceptance by potential users.
2. Sufficient demand for gas.
3. Ability to use the gas when produced.
4. Availability of sufficient raw materials to meet the production requirements.
5. Adequate maintenance and operational control.

ADVANTAGES OF BIODIGESTERS
__ Reduces organic content of waste materials by 30-50% and produces a stabilized
liquid effluent.
__ Provides a sanitary way for disposal of human and animal wastes.
__ Produces large amounts of methane gas which can be stored at ambient temperature.
__ Produces free flowing, almost clear liquid odorless effluent. The effluent is a good
liquid fertilizer.
__ Weed seeds are destroyed and pathogens are either destroyed or greatly reduced in
number.
__ Rodents and flies are not attracted to the end product of the process. Access of pests
and vermin to wastes is limited.
__ Helps to conserve on imported energy sources.

DISADVANTAGES OF BIODIGESTERS
__ Liquid sludge presents a potential water pollution problem if handled incorrectly.
__ Proper operating conditions must be maintained in the digester for maximum
gas production.(Guyana energy agency/Biodigester Manual)

CO2 scrubbing from biogas

A variety of processes are being used for removing CO2 from natural gas in petrochemical industries. Several basic
mechanisms are involved to achieve selective separation of gas constituents. These may include physical or
chemical absorption, adsorption on a solid surface, membrane separation, cryogenic separation and chemical
conversion. One of the easiest and cheapest method involves the use of pressurized water as an absorbent. The
raw biogas is compressed and fed into a packed bed column from bottom; pressurized water is sprayed from the
top. The absorption process is, thus a counter-current one. This dissolves CO2 as well as H2S in water, which are
collected at the bottom of the tower. The water could be recycled to the first scrubbing tower [3]. This perhaps is
the simplest method for scrubbing biogas. Savery et al. suggested that the three agents NaOH, KOH and Ca(OH)2
can be used in chemical scrubbing of biogas. The absorption of CO2 in alkaline solution is assisted by agitation. The
turbulence in the liquid aids to diffusion of the molecule in the body of liquid and extends the contact time
between the liquid and gas. Another factor governing the rate of absorption is concentration of the solution. The
rate of absorption is most rapid with NaOH at normality's of 2.5-3.0.

Gas purification can also be carried out using some form of silica, alumina, activated carbon or silicates, which are
also known as molecular sieves [12]. Adsorption is generally accomplished at high temperature and pressure. It has
good moisture removal capacities, simple in design and easy to operate. But it is a costly process with high
pressure drops and high heat requirements. Schomaker et al. [13] reported that CO2 could be removed from
biogas by pressure swing adsorption which consists of at least three active carbon beds. One of the beds is fed
with biogas under pressure (6 bar) CO2 is adsorbed. When there is saturation of CO2 in the adsorption bed, the
process is shifted to the second bed. The saturated bed is depressurized to ambient pressure. The efficiency of this
process is up to 98%.

Adsorption using iron oxide

H2S reacts with iron hydro-oxides or oxides to form iron sulfide. The biogas is passed through iron oxide pellets, to
remove H2S. When the pellets are completely covered with sulfur, these are removed from the tube for
regeneration of sulfur. It is a simple method butduring regeneration a lot of heat is released. Also the dust packing
contains a toxic component and the method is sensitive to high water content of biogas. Wood chips covered with
iron oxide have a somewhat larger surface to volume ratio than plain steel. Roughly 20 g of H2S can be bound per
100 g of iron oxide chips. The application of wood chips is very popular particularly in USA. It is a low cost product,
however, particular care has to be taken that the temperature does not rise too high while regenerating the iron
filter [9]. H2S can be adsorbed on activated carbon. The sulfur containing carbon can then either be replaced with
fresh activated carbon or regenerated. It is a catalytic reaction and carbon acts as a catalyst [15].

Liquid phase oxidation process

This process is primarily used for the treatment of gases containing relatively low concentration of H2S. It may be
either: (a) physical absorption process or (b) chemical absorption process. In physical absorption process the H2S
can be absorbed by the solvents. One of the solvent is water. But the consumption of water is very high for
absorption of small amount of H2S. If some chemicals like NaOH are added to water, the absorption process is
enhanced. It forms sodium sulfide or sodium hydrosulfide, which is not regenerated and poses problems of
disposal. Chemical absorption of H2S can take place with iron salt solutions like iron chloride. This method is
extremely effective in reducing high H2S levels. The process is based on the formation of insoluble precipitates.
FeCl3 can be added directly to the digester slurry. In small anaerobic digester system, this process is most suitable.
All other methods of H2S removal are suitable and economically viable for large-scale digesters. By this method
the final removal of H2S is about 10 ppm.

Biogas compression and storage

Biogas, containing mainly methane, could not be stored easily, as it does not liquefy under pressure at ambient
temperature (critical temperature and pressure required are - 82.5 8C and 47.5 bar, respectively).
Compressing the biogas reduces the storage requirements, concentrates energy content and increases pressure to
the level required overcoming resistance to gas flow. Compression is better in the scrubbed biogas. Most
commonly used biogas storage systems are given in Table 1 [18]. Integrated units with facilities for scrubbing,
compressing and storing have been developed in certain developed countries. For instance a water scrubber
coupled with a gas compressor is being promoted for uniform use in New Zealand. Similarly, the biogas produced
from poultry manure is being dried, scrubbed, compressed and stored at a pressure of 4 bar in 0.2 m3 steel tanks
in Belgium [19]. Khapre [6] conducted a study on scrubbing and compression of biogas and subsequently used it
for domestic cooking. He found reduced requirement of scrubbed and compressed biogas (0.353 m3) than raw
biogas (0.591 m3) for cooking a day's meal of a six member family. He stored the scrubbed and compressed biogas
at a pressure of 7 bar in cylinder of 0.1 m3 capacity. By purifying the biogas produced from the distillery wastes,
scientists of Jadhavpur University, Kolkatta, India [20] claimed to have generated huge quantities of compressed
methane, a gas with an immense potential and an alternative source of vehicle fuel.
Experimenting with bulk distillery wastes, from alcohol manufacturing breweries, researchers produced the gas by
bio-methanation of the effluents. Similar results have also been reported from Netherlands, UK, Australia, New
Zealand and USA. All these results indicate that biogas is one of the potential substitutes for present day fuels
including CNG, petrol, diesel and LPG [9]. Nema and Bhuchner [21] stressed on value addition to biogas by
scrubbing and compressing, making it as good as the compressed natural gas (CNG). They reported the economic
feasibility of producing energy from solid wastes of Delhi city. From 5000 tonnes wastes generated per day in
Delhi, 100,000 Nm3/day biogas can be produced which is equivalent to 309.5 m3 CNG worth US $ 70,000 per day.
Beside this, by adopting this technology 117 tonnes/day CO2 gas can be prevented from entering into the
atmosphere.

Biogas scrubbing, compression and storage:

perspective and prospectus in Indian context
S.S. Kapdi, V.K. Vijay*, S.K. Rajesh, Rajendra (Pure Biogas)

The Brief
The purpose of this project was to design and build an anaerobic digester to meet the following criteria.
The design should
__attempt to maximize the amount of biogas produced per unit time,
__be a continuous flow anaerobic digester. This has been specified because it seems that this will be the most
practical design for continuous operation in a farm situation.
__be simple and easy to understand so that the average person is able to grasp the function and theory behind
each
component of the design with only a small amount of guidance. The idea here is to encourage people looking at
the design to think and understand the requirements for controlled anaerobic digestion and the continuous flow
model.
__be a durable, compact, versatile design which is capable of being shifted around if necessary to be displayed.
__be operated with a minimum of monitoring, regulating, and adjusting (in other words, be easy to operate).
__attempt to reduce time and money costs associated with maintenance

__attempt to minimize the cost of setting up and running the digester without compromising the performance of
operation or the other specifications of the brief
__look aesthetically pleasing as another mechanism to effectively sell the concept!
The Design and Theory of a Basic Anaerobic Digester by Simon Knowles(Basic Biogas Digester)

Picture. 1. Scheme of biological transformation organic compounds under anaerobic

conditions.
As a result of biological decomposition proteins, lipids, carbohydrates are obtained:
Organic acids acetic, butiric, propionic, formic, caproic, lactic;
Alcohols and cetones methanol, ethanol, isopropyl, glycerol, butanol, acetone;
Gases methane, carbon dioxide, hydrogen, ammonia, hydrogen sulfur;
Enzymes - cellulase;
Vitamins riboflavin (2), cyancobolamine (12).
These products are intermediates and can be found inside bioreactor.
Final products of anaerobic biodegradation of organic compounds are:
biogas (methane content > 55%, carbon dioxide < 45%, hydrogen sulfur < 2%,
hydrogen < 1%);
fermented substrate containing water, cellulose residues, small quantity of bacteria
biomass and inorganic substances (nitrogen, phosphorus, potassium, sulfur and
others).
Technology process division is made accordingly to microbial conversion stages. In the first bioreactor hydrolysis
and partial acidogenesis of organic substances occur. In the second one acidogenesis completes and acetic acid,
carbon dioxide and methane begin to form. The two stage scheme comparing to one stage, helps better control
organic substances conversion and receive stable biogas formation with high efficiency and low time and energy
loses. Many companies implement simpler one stage technology. But two stage technology more economical.
Under the same conditions of digestion time and energy consumption at one stage technology lower biogas
volume can be produced. And otherwise for the same biogas yield at two stages scheme needs less time and
energy. Full mechanic agitation at one stage needs twice more agitators and electricity than at two stage scheme.
At organic wastes quality deviation biogas yield sharply decrease at one stage technology while two stage
technology much more stable and allow to regulate biogas production at both stages.
Fermented substrate is reasonably to separate onto two fractions liquid (filtrate) and solid
(fugate).
Fugate is ready for use organic fertilizer (pic.2). Filtrate can be used in different ways for dilution of incoming
wastes to the necessary water content, nourishing plants at fields or after its aerobic cleaning to the allowed level
releasing to natural basins.

DESCRIPTION
to commercial offer
a biogas plant for conversion of organic wastes to biogas
Cleaning Recycling & Organic Fertilizer(Biogas Plant Food Waste)

The benefits of biogas generators are explicitly listed below and should be made clear when suggesting the
construction of the biogas generator to users in order to improve speed and likelihood of acceptance:
1.Biogas generators provide a safe and cleaner wayof storing excreta and subsequently bring aboutrelated
advantages linked to safe sanitation
2.Biogas generators provide free fuel for cooking,heating and lighting
3.Biogas generators provide fertiliser for crops
4.Biogas requires far less time and effort to collectthan other fuels (e.g. wood)
5.Biogas reduces the need for wood and thereforereduces deforestation and the burden on women ofcollecting
wood
6.Biogas creates no smoke and therefore reduceshealth problems caused by burning other fuelsindoors
7.Biogas is environmentally friendly and does notrelease as many greenhouse gases when burnedcompared to
other fuels
8.Dangerous bacteria in faeces are killed duringdigestion in the biogas generator

Biogas use
Appliances connected to the biogas distribution pipes can extract gas from the gas holder when required.
Cooking Stoves specifically designed for biogas are available which make the best use of the fuel a much higher
fuel:air ratio is required with biogas burners compared to butane burners (1:6 by vol. compared to 1:30)[5]. These
are available from a number of manufacturers for between US $15-$30 (e.g. Rupak Enterprises, India) [9]or can be
made more cheaply locally (see Case Study II).
Alternatively simple modifications can be made to butane/propane stoves (by expanding the gas jet cross-section
by 2-4 times or modifying fuel:air ratio by adjustment if possible) to obtain the desired compact and slightly
blueish flame
Process parameters
There are a number of parameters which effect the production of biogas which should be kept at an optimum level
for human excreta biogas generators:
1. Substrate temperature Digestion works best at around 36c. Expected approximate gas return of 1kg of
human waste over 60 days is as follows:

o 0.43m3 biogas at 35c

o 0.3m3 biogas at 25c

o Unsatisfactory under 15c

2. Hydraulic retention time (HRT) The average amount of time that the liquid part of the slurry shall be in the
digester for. This should be long enough to reduce many of the pathogens and to allow the maximum amount of
gas to be extracted however will increase the digester volume. The HRT should be based on a compromise of
pathogen removal time and digester size. Commonly the HRT lies in the range 60-100 days if no wastewater
sewage outlet is available however can be reduced to as little as 5 days [8] in the presence of a sewage outlet. A
summary of pathogen inactivation times (within the sludge) is shown in Table 5:

3. Solid retention time (SRT) The sludge (more solid part of substrate) should be stored for a relatively long
retention time (anywhere between 1 and 5 years depending on digester design, waste content, digester size, etc)
before being removed from the generator and pre-treated for the appropriate time prior to use as fertiliser (e.g.
high

temperature composting with added soil components, see Table 7). Over this period of time the maximum amount
of gas will have been produced and the number of pathogens in the waste should have decreased considerably.
The solid retention time is very much variable and the ideal is generally found by experiment and experience.

4. pH The pH of the slurry should not drop below 6.2 (this will have a toxic effect on methane-producing
bacteria). A healthy digestion process is indicated by a neutral pH (7.0) [6].
5. Agitation/mixing Mixing of the slurry can increase gas production by ensuring an even distribution of bacteria
and fresh substrate. The large scale industrial generators are often fitted with motor-driven rotating paddles whilst
smaller agricultural ones are mixed with long poles by hand. There is little information on the optimum frequency
of mixing in human waste fed digesters but GTZ suggest the gas production increases dramatically when mixing is
undertaken (slowly and perhaps once a day or once a week). Different time intervals between mixes should be
tried to identify the optimum level for any specific generator.

6. Solids content Generally a slurry with a solids:liquids (faeces:urine) ratio of 1:1 should be aimed for. A Total
Solids (TS) content of between 7- 11% is ideal since the actual liquid content of faeces is quite high faeces and
urine should be added in approximately equal amounts to achieve this.
7. Inhibiting factors The presence of heavy metals, antibiotics and detergents in the slurry can inhibit the biogas
production process. Any addition of these to the digester should be avoided. Anything which is not biodegradable
should not be added to the digester since it will take up valuable space and could lead to a blockage. It would seem
that anal cleansing material (including water and paper) can be added to the digester [19] whilst keeping in mind
that the ideal solid:liquid ratio of 1:1 should be adhered to where possible.

8. Carbon:Nitrogen ratio The carbon:nitrogen ratio of the inlet waste should be in the region 9-25:1 for efficient
biogas production (the methane producing bacteria work well with this ratio). Degradable food, agricultural and
animal waste can be mixed to the correct solid:liquid ratio (1:1) in an influent collecting

tank prior to addition to the main digester. Different types of waste can be added to alter the C/N ratio to reach an
ideal. Approximate values for C/N ratios and TS values are shown in Table 6.
9.Location To increase the temperature of thesubstrate, sunny locations (i.e. away from trees orshade) are good
locations for biogas generators.As a precaution for (unintended) leaching,generators should be situated at least
30m awayfrom any water source or stream. No permanentstructures or through ways should be build on
theground above a generator.

10.Fertiliser output Per day an average adult willexcrete around 10-12g of nitrogen, 2g ofphospherous and 3g of
potassium sludge/slurryfrom the generator has excellent potential for useas agricultural fertiliser.
Since many biogas generators run on a continuouscycle it is necessary to post-treat any sludge priorto use as
fertiliser (retention times in continuousgenerators cannot be guaranteed). A number ofpost-treatment options are
suggested in Table 7.
VD = VB x HRT (Eq. 1)
Where:
VD = Volume of the digester (m3)
VB = Volume of biomass added per day (m3/day)
HRT = Retention time required (days)

The amount of human waste produced varies from person to person but generally lies in the region of 0.2-0.4kg
(solid) and 1-1.3kg (liquid) per day (depending on diet, health, etc). If other waste (animal dung, organic food
waste, etc) is added then this should also be taken into account. Clearly it is almost impossible to control the rates
of waste input (especially in the case of latrines) so some discretion and common sense should be used when
dealing with the numbers.

The volume of the gas holder VG depends on the relative rates of gas production and consumption. To calculate
the daily gas production (G) either Equation 2 or Equation 3 can be used (it may be good to use both and take an
average since data for Gy varies greatly):
oG = VB x Gy(moist mass) (Eq. 2)
oG = LSU x Gy(species)(Eq. 3)

Where:
G = Daily gas production rate (m3/day)
MB = Mass of biomass added per day (kg/day)
LSU = Number of live stock units (number)
Gy(moist mass) = Gas yield per kg of excreta per day (m3/kg/day)
Gy(species) = Gas yield per kg of live stock unit per day (m3/kg/day)

The gas holder must be designed to cover the peak consumption rate (VG1) (if the primary reason for construction
is based on biogas demand) and the longest period of zero consumption (VG2) (if the primary reason for
construction is safe excreta treatment/disposal). The larger of these 2 volumes should be used to specify the gas
holder volume with an additional 20% safety margin. The following equations should be used to calculate VG1 and
VG2:
VG1 = Gcmax x Tcmax (Eq. 4)
VG2 = G x Tczero (Eq. 5)
Where:
VG1 = Gas holder volume 1 (m3)
VG2 = Gas holder volume 2 (m3)
Gcmax = Maximum rate of gas consumption (m3/day)
Tcmax = Maximum time of gas consumption (days)
G = Daily gas production rate (m3/day)
Tczero = Maximum time of zero gas consumption (days)
The final gas holder volume can then be calculated using the largest of VG1 and VG2 with Equation 6 below:
VG = VGmax + (VGmax x 0.2) (Eq. 6)
According to GTZ (from experience) the ratio of digester volume:gas holder volume (i.e. VD:VG ) usually lies in the
range 3-10:1.
Design, construction and maintenance of a biogas generator(Design construction and maintenance of a biogas
generator)

There are four types of methano-genic bacteria; Methano-bacterium, Methanospirillium, Methano-coccus and
Methano-circina. These bacteria are oxygen sensitive and photosensitive and do not perform effectively in the
presence of oxygen and light. The gas thus produced by the above process in a bio-gas plant does not contain pure
methane and has several impurities. A typical composition of such gas obtained from the process is as follows:
Methane 60.0%
Carbondioxide 38.0%
Nitrogen 0.8% Hydrogen 0.7%
Carbon-monoxide 0.2%
Oxygen 0.1%
 Hydrogen Sulphide 0.2%

The benefits derived from biogas plants in terms of manure and useful energy are environment friendly. The
average NPK content of Farm Yard Manure (FYM) is about 0.5, 0.2 and 0.5 percent respectively and it may be
observed that biogas slurry is rich in NPK by more than four times than ordinary dung when converted into FYM.
When the country is faced with shortage of fertilizers and has to spend enormous amounts for its import, the
application of bio-gas slurry can replace the chemical fertilizers to a large extent. Bio-gas slurry or FYM not only
adds NPK but it proves the soil porosity and texture. These are established benefits. Second major benefit is that
rural people would gradually stop felling trees. Tree felling bas been identified as one of the major causes of soil
erosion and worsening flood situation. Government has started massive afforestation programme to tackle the
erosion and flood situation. Continued deforestation has been causing ecological imbalances in the environment in
which we live. Bio-gas plants would be helpful in correcting this situation. In rural areas kerosene is used for
lighting lantern and cooking in a limited way wherever kerosene supply has been made possible. Whatever
quantity is used can be replaced by bio-gas as it can be used for lighting and cooking. This would reduce the
dependence on fossil oil directly and in saving foreign exchange.
The GHG potential of methane is higher than of carbon dioxide by 23 fold and of nitrous oxide by 296 fold. When
biogas displaces fossil fuels from energy production and transport, a reduction of emissions of CO2, CH4 and N2O
will occur, contributing to mitigate global warming. The thing to understand is this, Methane gas is produced by
Methanogens, and Methanogens prefer fatty acid chains that are 1-6 carbons in length, they prefer simple foods,
and not complicated ones. These fatty acids are known as Volatile Fatty acids.
For this to occur, in enter ourtrusty bacteria friends.(ICESN/ Biogas directory)
It can be seen now why compression alone will not liquefy methane. However, methane can be compressedto a
gss volume so small as to be comparablet o a liquefied gas as far as volume reduction is concerned, but certain
problems arise: 1) the tank container must be extra strong, thick, heavy and of special construction; 2) a costly high
compressor systemi s necessary;3 ) expensiveg as pressurer eduction devicesa re neededf or every tank
when the compressedg as is used

There 8fe many advantages to be gained in transporting methane as a liquid even with the smm requirement of
maintaining a temperature of at most - 82OC and a pressure of at least 45.8 atmospheres throughout the time of
transport and storage. The volume of the gqu& however, is approximately only l/600& that of the gas at 25OC and
this is an advantage in storage. The problems met in handling gases at such cryogenic temperatures have been
quite satisfactorily solved in the development of space rockets.
The Bacteria in Biogas Production Pig manure, by itself, generates biogas spontaneously, although it takes quite
some time for this to happen. Hobson and Shaw (1967), using a M-liter digester initially filled with water, added
pig manure gradually so that the water was replaced in about 4 weeks time. By then the total solids was 2%.
Operating as a continuous-fed system, pig manure was added daily at the rate of 0.03 lb. per cu. ft. per day. At
regular time intervals they determined the kind and number of bacteria. Their results were as follows: First week:
Total count was 5 x lo6 to 5 x 10 /ml. The number of amylolytic bacteria (starch decomposers) was greater than 4
x 1oQnl. Third week: By this time the methanogenic bacteria (methane producers) began to appear, numbering
about lo3 /ml. and they increased in number with increasing time.
Fourth to fifth weeks: The cellulolytic bacteria (cellulose decomposing), numbered lo4 to 10 5 /ml. The proteolytic
bacteria (protein-decomposing) also appeared at about this time at greater than 4 x lo4 /ml. Ninth week: The
methanogenic bacteria reached 10 /ml.
The nonmethanogenic bacteria were found to be principally the noncellulolytic and
the cellulolytic bacteria.
I. Noncellulolytic bacteria.
1. Streptococci, facultatively anaerobic, constituting 43 to 47% of all isolates. Nonproteolytic
and nonamylolytic, probably play a role in maintaining the anaerobit
condition in digesters.
16
: 2. Bact&ds, constituting 20 to 80% of the anaerobic bacteria. Gram-negative
pleomorphic rods, short to medium length; some are coccobacilli, mostly amylolytic.
Fcrment mono- and disaccharidesa a well as glycerol producing propionic,
acetic and butyric acids.
3. Clostridia. The most active proteolytic bacteria,found belong to three groups:
A. Amylolytic; very similar to Clostridium butyricum; fermentation products
are acetic and butyric acids.
B. Proteolytic; ferment sugars forming acetic and isovaleric acids.
C. Proteolytic but do not ferment sugar.

The methanogenic bacteria possess the following general characteristics (Barker,

1956):
1. Strictly anaerobic; not only molecular oxygen but also compounds that easily
give oxygen like the nitrates must be absent. In Barkers studies (1956), pure
cultures of these bacteria were obtained successfully for the first time only by
using sodium sulfide to remove the last traces of oxygen.
2. Require a pH range for growth of 6.4 to 7.2 (other authors put the optimum
pH at 7.2 to 8.2). However, one species grows at pH 8-9, and in peat bogs where
the pH is about 4, some methane is also formed.
3. Utilize ammonium salts as nitrogen source.
4. No known need for nutritional factors (probably amply supplied by commonly
used substrates). This is in contrast to other organisms like yeast.
5. Produce methane as a major metabolite.
6. Exhibit extreme substrate specificity; these bacteria are able to utilize only a few
very simple compounds

In accordance with known microbiological techniques, attempts have been made

to grow in strict isolation, each species of bacterium that produces methane. This has
proven to be very difficult; hence there are only very few that are definitely confvmed
to be methane producers.

Barker( 1956)c lassifiedth e methanogenibca cteriaa s fdlows:

Family: Methanobacteriaceae
A. Rod-shapedc ells
i. Nonsporulatiqg Methunobucterfum
1. Mikuxfonnicfcum -------- CO, H, , formate-/
2. Mkt. propionicum ---- propionate
3. M&act. sohngenii ----------------- acetate, butyrate
4. M&t. ruminantiud~ -------- CO2 , H 2
II. Sporulating: Methanobacillus
1. Mbac. omelianskii ----------------- H.2 , primary and secondary alcohols
B. Spherical cells
I. Cells not in sarcina arrangement: Methunococcus
1 l MC* m& - ~~~~~~~~~~~~~~~~~~~ - ---- ----- acetate, butyrate
2. MC. vannielif ---- - ------ ---------------- forma@, H 2
II. Cells in sarcina arrangement: Methanosurcinu
1 a Ms. bark&i --------------------------- CH3 OH, acetate, CO, H2
2. Ms. methanica ------------------------ acetate, butyrate
Methane Production in the Rumen - Pertinent to this discussion on methane producers
are the results of studies on the microflora of the rumen. Hungate and co-workers
(through Thimann, 1963) have shown that the rumen contains many organisms closely
related physiologically, mostly short rod, oval or coccus, which actively ferment cellulose
to organic acids like acetic and propionic. Methane results from the secondary
fermentation of these acids by methanogenic bacteria which have been found present up
to 2 x 10 /ml. in rumen fluid. A cow reportedly gives as much as 700 liters of gas
in a day.
(Biogas and waste recycling: the Philippine experience/Biogas Philippines)

Temperature
The optimum temperature, i.e. the temperature at which the organism grows fastest and works most efficiently,
varies among species. Microorganisms can be divided into different groups depending on the temperature at
which they best thrive and grow: psychrophilic, mesophilic, thermophilic, and extremophilic/hyperthermophilic
(Noha and Wiegel 2008). Typically, the optimum temperature for a specific organism is strongly linked to the
environment from which it originates. For example, microorganisms that live in marshland, tundra, or in a septic
tank, may have a low optimum temperature (around 10C) (psychrophilic temperature range), whereas human
intestinal bacteria, such as Esherichia coli, grow best at 37C (mesophilic temperature range).

Organisms with an optimum temperature above 50C are called thermophiles, and those that grow above 65C are
called extreme thermophiles (Noha and Wiegel 2008). Some microbial communities are adapted to grow at even
higher temperatures. Microorganisms that have very high optimum growth temperatures (above 85C) live in hot
springs and submarine volcanoes. The latter belong to the so-called hyperthermophiles, in which the cell proteins
and other components are intact even at these high temperatures (Wagner and Wiegel 2008).
Common to all growth intervals is that the temperature that allows the highest rate is close to the so-called
maximum temperature, which results in cell death. If the temperature increases above this maximum
temperature, the cell's proteins and other components are quickly inactivated, causing the organism to die. The
maximum temperature varies depending on which temperature range the microorganism is adapted to.

A biogas process contains many different organisms, and to some extent, they differ in how they respond to
temperature. However, the biogas process usually operates at a temperature range of around 30C-40C or 50C-
60C (Nordberg 2006). Biogas production is possible at psychrophilic temperatures but may also result in a lower
methane production rate depending on the type of process (Hesselgren et al 2005, Collins et al 2006, Bohn et al
2006). In the case of high temperatures, there are examples of methane-producing organisms that can handle
110C (Chaban et al 2006), but stable biogas processes do not seem to operate above 60C-70C (Scherer et al
2000). At temperatures above 60C, the activity of methane producers is reduced to a greater degree than that of
acid-forming organisms, which often results in the accumulation of fatty acids in the biogas process (Nozhevnikova
et al 1999, Scherer et al 2000).

Oxygen
The importance of oxygen concentration varies greatly for the different microbial communities that comprise the
biogas process. Some of the organisms, such as those that produce methane, are very sensitive to oxygen and die
if they come in contact with air. Others can survive quite low concentrations of oxygen, while others grow better if
oxygen is present. The free radicals of oxygen are strong oxidising agents that can destroy cells by oxidizing various
cell components. Microorganisms that can live in the presence of oxygen have different defence systems, that is,
various enzymes that can protect the cell against oxidation by oxygen. The organisms that are sensitive to oxygen
do not have this enzymatic defence system and are destroyed in the presence of air. Microorganisms are usually
divided into different groups depending on their relationship with oxygen. Both strict anaerobes and so-called
facultative aerobes are found in the biogas process. Strict anaerobes only grow in the absence of oxygen. This
group includes the methane-producing organisms. On the other hand, facultative aerobes grow in both the
presence and absence of oxygen. This group includes numerous fermentative microorganisms. In the presence of
oxygen, they can grow by aerobic respiration, but then they switch to fermentation when oxygen is depleted. This
means that a temporary air leakage to a biogas process need not be a problem because there are microorganisms
that can rapidly consume the incoming oxygen. There are even studies that show that a brief aeration during the
biogas process can be a way of reducing the concentration of fatty acids (Agdag and Sponza 2004).

pH
Most microorganisms prefer a neutral pH range, i.e. about pH 7.0-7.5. However, some organisms are active at both
lower and higher pH values. There are several different organisms in the biogas process, and their pH requirements
for optimal growth vary greatly. While fermenting, acid-producing microorganisms manage to live in relatively
acidic conditions, down to pH 5.0, most methane producers generally require neutral pH values to be active.
Although most methane producers thrive best at neutral pH values, they remain active outside this pH-range
(Whitman et al 2006). There are known examples of acidophilic methane producers that grow down to pH 4.7
(Bruer et al 2006) and alkaliphilic methane producers that grow at pH values of up to 10 (Mathrani et al 1988).
Several biogas processes are currently operating in Sweden at pH values around 8 (personal communication,
Anders Ek, Swedish Biogas) and the literature also contains examples of biogas processes operating at a pH values
below 6 (Savant et al 2002). The fact that acid-forming organisms can handle a lower pH is illustrated by the fact
that decomposition of the substrate often begins already in the substrate tank, with acid formation and low pH as
a result. However, methane production does not usually occur here because the pH is too low. Instead, it starts in
the digestion tank where the pH is significantly higher. The growth of microorganisms at various pH ranges often
follows the same pattern as the growth at various temperatures. That is, at all growth intervals, the pH value that
generally results in the greatest rate is closest to the pH value that results in cell death.

Salts
All microorganisms require salts to function. The salts contain essential building blocks for the microorganisms,
such as sodium, potassium, and chlorine. These substances are available in many substrates and do not need to be
added to the biogas process separately. However, some waste has a high salt concentration or results in the
release of excess salt, which can inhibit the microorganisms in the biogas process. Salts (and sugars) generally have
a preservative effect, that is, they inhibit bacterial growth. Too much salt (or sugar) causes the cell to pump out
water and lose both form and function. Some organisms can adapt to high salt concentrations if they are allowed
to adjust slowly. They often form so-called osmolytes: compounds that help them maintain their function, even in
the presence of salt. Organisms that can handle relatively high salt concentrations are called halotolerant, and
those that grow even better at high salt concentrations are called halophiles. The most extreme forms of halophile
grow best at salt concentrations above 20%-30% sodium chloride (> 3.4mol/L-5.1mol/L) and this group also
includes some methane producers (Chaban et al 2006). Examples of materials that could lead to increasing salt
concentrations in biogas processes are waste from the food and fisheries industries, or different types of protein-
rich materials that lead to the release of ammonia. Typically, methane-producing microorganisms are usually the
ones most affected by increasing salt concentrations in a biogas process (see Chapter 4).

What temperature should be selected?

Mesophilic or thermophilic: which process is preferable? Generally, the process is faster at higher temperatures.
The heat makes the microorganisms work faster, provided that the species being added are adapted to this
climate. More of the materials is broken down in less time and the digestion tank volume can be reduced
compared to if the same amount of material were digested at a lower temperature (Duran and Speece 1997,
Edstrm and Nordberg 2004). A highertemperature may also increase the availability of certain organic
compounds because solubility generally increases with increasing temperature. As a result of increased solubility,
the viscosity of certain materials may be lower in thermophilic conditions, which facilitates mixing (van Lier 1995,
Ryan 2008). Another advantage of thermophilic digestion is that the high temperature provides natural sanitation
of the material; undesired pathogenic microorganisms such as Salmonella are destroyed more efficiently at higher
temperatures (Sahlstrm 2003). The time that the material is exposed to the high temperature is critical for
whether it is possible to achieve adequate sanitation.

Loading
Biological decomposition of organic matter occurs continuously in a biogas process. If no new material is added,
the process gradually stops. Loading is a term that indicates how much new material is added to the process per
unit of time. It is usually referred to as organic loading or organic loading rate (OLR). In this case it is important to
know the dry solids (DS) and organic matter (VS [volatile solids]) content in the substrate in order to give the
biogas process the right loading rate. Dry solids are the material that remains when all of the water is dried off,
while VS indicates the organic part of the dry solids.

Retention Time
Retention time is defined as the time it takes to replace all of the material (entire volume) in the digestion tank. In
a biogas process, hydrocarbons in solid form are converted to methane gas and carbon dioxide gas. Thus, the
amount of solid material is continuously reduced, i.e. the content of organic matter decreases during the process.
When new carbon-containing substrate is added to the process, this will also be converted to gas. Typically,
however, more substrate is added than is completely decomposed between each addition of substrate. By
removing part of the contents of the digestion tank at regular intervals, a constant volume is maintained in the
process. The volume of solid material being added is sometimes greater than the volume of solid material removal,
because some is removed as gas during the process. The volume of added and removed material is also regulated
by the amount of liquid added. The removed material is partly composed of water, including dissolved salts, and
organic matter that has been digested in the chamber for shorter or longer periods of time, known as digestion
residue. This residue also contains biomass, i.e. microorganisms grown during the process. The removed material,
e.g. water, dissolved salts, digestion residue and biomass, is often termed digestate.

Hydraulic and Solids Retention Time

Retention time is usually referred to as hydraulic retention time (HRT), and for the biogas process it is usually
between about 10 and 25 days, but can also be longer. Sometimes the retention time of the particulate material,
or solids retention time (SRT), in the process is listed instead. In many cases, HRT and SRT are equal, but in a
digestion tank in which part of the residues are returned to the process, SRT becomes longer than HRT. This may
occur, for example, during digestion of industrial sewage sludge, where added material has a high water content
and where the recirculation of digested, thickened sludge, including biomass, allows a longer time for the
microorganisms to break down the incoming organic matter.
The length of the retention time needed, depends partly on the composition of the substrate and the digestion
temperature. Microorganisms generally manage to decompose a substrate rich in sugar and starch, which is easily
broken down, in a short time. An example is industrial waste water that only contains soluble organic matter. In
this case, no hydrolysis is necessary, which allows for a relatively short HRT. On the other hand, microorganisms
may need significantly more time to effectively attack and break down fibre-rich and cellulose-rich plant matter.
For such material, it is often hydrolysis and not methanogenesis that limits the rate of decomposition. In Germany,
among other places, retention times of up to 50-100 days are used to ensure stable operation and satisfactory
digestion of energy crops (Ergebnisse des Biogas Messprogramms 2005).

Microorganisms can be retained in the process

If the retention time is much too short, there is a great risk that the microorganisms will not manage to grow at the
rate at which material is removed from the process. As mentioned earlier (Chapter 1), the dominant methane
producers in a biogas process, often have doubling times of up to 12 days. This means that the retention time for
the material in the digestion tank can seldom be shorter than this. In other cases, the microorganisms are simply
rinsed out in such large numbers that the populations do not have time to recover until the next time digested
matter is removed. Thickening the sludge before digestion increases the dry solids content and the concentration
of active microorganisms in the digestion tank. This method is often used in Swedish waste water treatment plants
(VAV P42 1981).

Mixing
Digestion tanks should be equipped with agitators (mechanical agitators or pumps) to mix the substrate. Mixing
facilitates contact between the microorganisms, the substrate and nutrients and provides a uniform temperature
throughout the process. It is particularly important for hydrolytic microorganisms to make good contact with the
various molecules that they should digest and that their enzymes can be distributed across a large surface area
within the substrate. Mixing also prevents material from accumulating on the bottom of the digestion tank and
reduces the risk of foaming.
Mixing also facilitates the important contact and transfer of hydrogen between methane producers and the
organisms that carry out anaerobic oxidation. However, mixing ought not to be too strong. Often these
microorganisms grow in tight clumps, called aggregates, which facilitates their close cooperation and thus the
transfer of hydrogen. Gentle mixing benefits the formation of aggregates and prevents methane producers from
being washed out in the liquid. Continuous mixing avoids sedimentation and utilises the existing digestion tank
volume in the best manner. Substrates with high dry solids content are generally harder to mix than more liquid
materials. Mixing in the substrate tank is also important to avoid sedimentation and thus uneven loading of the
digestion tank.

Alkalinity and pH
Biogas processes usually run best at neutral pH values or slightly above neutral (pH between 7.0 and 8.5).
Maintaining neutral and stable pH values requires that the process has a relatively high and constant alkalinity.
Alkalinity is a measure of the amount of alkaline (basic) substances in the biogas process. The higher the alkalinity,
the greater the buffer capacity in the process, which in turn promotes a stable pH value. Alkalinity consists
primarily of bicarbonate ions that are in equilibrium with carbon dioxide (Equation 1). Carbon dioxide and
carbonate ions also contribute to alkalinity. Decomposition of nitrogen-rich substrate with high proportions of
proteins and amino acids can increase alkalinity, because the ammonia released can react with dissolved carbon
dioxide to form ammonium bicarbonate.