CHAPTER-1

INTRODUCTION
1.1 General:
Anaerobic digestion is a collection of processes by which microorganisms break
down biodegradable material in the absence of oxygen. The process is used for
industrial or domestic purposes to manage waste or to produce fuels. Much of
the fermentation used industrially to produce food and drink products, as well as
home fermentation, uses anaerobic digestion. This happens in a oxygen free sealed
tank called Anaerobic digester.
Anaerobic biological treatment is well understood and used frequently as anaerobic
digesters to treat complex organic solid wastes such as primary and secondary
wastewater sludges. However, it has not been used much in the past to treat low
strength organic wastewaters from industrial and domestic applications. Aerobic
processes were preferred for treatment of these wastewater streams because they are
easy to operate and can tolerate process fluctuations. In comparison, anaerobic
reactors were assumed to be less stable under fluctuations, more expensive to install
and require long start-up time. This belief was due to limited knowledge of the
process and reactor design. Now the technology advances have significantly reduced
the historical weakness of anaerobic treatment. With the work of Young and McCarty
in the year 1969, application of anaerobic process for the treatment of industrial and
municipal wastewaters has gradually increased in last three decades. Today the
anaerobic treatment has emerged as a practical and economical alternative to aerobic
treatment due to significant advantages over aerobic treatment.
1.2 Main objectives:
 To present about the working principle and the anaerobic digestion
process
 To present about the different types of anaerobic digesters
 To present about the factors influencing the anaerobic digestion process
 To explain about the merits and demerits of anaerobic digestion process
 To explain about the applications and limitations of the anaerobic
treatment process

1
 CHAPTER-2
LITERATURE REVIEW
2.1 Anaerobic digestion technology in poultry and livestock waste treatment
Suleyman sakar , kaan yetilmezsoy,emel kocak
A literature review has been undertaken to investigate the performance of the different
anaerobic process configurations and operational conditions used in poultry and
livestock waste treatment. The results of the extensive literature review showed that a
wide range of different reactor volumes varying from 100 mL to 95 m  3 were utilized
in the investigation of anaerobic processing of poultry manure. Retention times
studied were between 13.2 h and 91 days. Most of studies were carried out under
mesophilic conditions maintained between 25o and 35°C. Chemical oxygen demand
(COD) removals and organic loading rate (OLR) ranged from 32 % to 78%, and from
1.1 kg to 2.9 kg COD m -3 day-1, respectively. Biogas yields were achieved between
180 mL g-1 COD added and 74 m3 day-1 for a wide range of different reactor
configurations. Up-flow anaerobic sludge blanket (UASB) seems to be a suitable
process for the treatment of poultry manure wastewater and the liquid fraction of hen
manure, due to its ability to maintain a sufficient amount of active biomass. The
literature review showed that various reactor configurations such as fixed-film reactor,
attached-film bioreactor, anaerobic rotating biological reactor, batch reactors,
downflow anaerobic filter, fixed dome plant, UASB, continuously stirred tank reactor
(CSTR), up-flow anaerobic filter (UAF), temperature-phased anaerobic digestion
(TPAD), anaerobic hybrid reactor (AHR), and two-stage anaerobic systems are well
suited to anaerobic processing of cattle manure. At both mesophilic and thermophilic
conditions, high COD removals (87%-95%) were achieved for treatment of cattle
manure wastewaters. The COD and volatile solids (VS) reductions obtained were 37.9
to 94% and 9.6 % to 92%, respectively. During the studies, OLR and retention times
ranged between 0.117 and 7.3 g VS L -1 day-1 and between 0.5 days and 140 days,
respectively. In anaerobic processing of cattle manure, methane yields between 48
mmol CH4 L-1 and 4681.3 m3 CH4 month-1 were found for the corresponding reactor
volumes of 120 mL and 1300 m3, respectively. In anaerobic processing of swine
manure, OLR ranged from 0.9 to 15.42 g VS L-1 day- 1 at mesophilic conditions (25o-
35°C). The reactor volumes varied between 125 mL and 380 L. Temperature and
retention times ranged from 25oC to 60°C, and 0.9 to 113 days, respectively. COD

2
and VS reductions achieved were between 57% and 78% and between 34.5% and
61%, respectively. Moreover, methane yields were obtained between 22 mL and 360
mL CH4 g - 1VS added. The results showed that UASB, anaerobic baffled reactors,
CSTR, and anaerobic sequencing batch reactor (ASBR) were successfully utilized in
anaerobic processing of swine manure at both mesophilic and thermophilic
conditions.
Keywords :Poultry manure, livestock manure, anaerobic digestion, hydraulic
retention time, biogas, wmr 1167-6

2.2 Water hyacinths as a resource in agriculture and energy production

Carina c.Gunnarsson

Water hyacinths are becoming a problem in lakes, ponds and waterways in many parts
of the world. This paper contains a literature study of different ways to use water
hyacinths, mainly in agricultural or alternative energy systems.

The literature review indicated that water hyacinths can be rich in nitrogen, up to
3.2% of DM and have a C/N ratio around 15. The water hyacinth can be used as a
substrate for compost or biogas production. The sludge from the biogas process
contains almost all of the nutrients of the substrate and can be used as a fertiliser. The
use of water hyacinth compost on different crops has resulted in improved yields. The
high protein content makes the water hyacinth possible to use as fodder for cows,
goats, sheep and chickens. Water hyacinth, due to its abundant growth and high
concentrations of nutrients, has a great potential as fertiliser for the nutrient deficient
soils of Africa and as feed for livestock.

Applying the water hyacinths directly without any other processing than sun drying,
seems to be the best alternative in small-scale use due to the relatively small losses of
nutrients and workload required. To meet the ever-growing energy demand, biogas
production could be one option but it requires investments and technological skills
that would impose great problems in developing countries where the water hyacinth is
often found. Composting as an alternative treatment has the advantage of a product
that is easy to work into the soil compared with dried water hyacinths, because of the
decomposed structure. Harvesting and transport of water hyacinths can be conducted
manually on a small scale and does not require a new harvesting technique to be
introduced. Transporting of fresh water hyacinths means, if used as fertiliser in
3
amounts large enough to enhance or effect crop growth, an unreasonably large labour
requirement. Based on the labour need and the limited access to technology, using
dried water hyacinths, as green manure is a feasible alternative in many developing
countries.

2.3 Kinetics of Anaerobic Treatment 

S. G. Pavlostathis, E. Giraldo-Gomez

A review of the kinetics of anaerobic treatment and the reported values of such kinetic
parameters as the maximum specific substrate utilization rate (k), the half-saturation
constant (Ks), the microbial growth yield (Y), and the microorganism decay rate
constant (b) are presented. The available kinetic information is presented for each
subprocess: (a) hydrolysis of complex, paniculate organic materials; (b) fermentation
of amino acids and sugars; (c) anaerobic oxidation of long-chain fatty acids and
alcohols; (d) anaerobic oxidation of intermediary products (such as short-chain fatty
acids); (e) homoacetogenesis; and (f) methanogenesis. The intrinsic rates of each step
as well as mass transfer limitations and their effect on the intrinsic kinetics are
discussed and areas requiring further research are also identified. Substantial variation
exists in the reported values of the kinetic coefficients. This variation is due in part to
the variability in mode of operation, environmental and operational conditions in the
various studies as well as to the lack of a widely accepted standard procedure for
measuring and expressing the biokinetic coefficients. The hydrolysis step is usually
assumed to follow first-order kinetics. Whenever the kinetics of the hydrolysis step
were studied, they were generally found to be the limiting-step in the overall
conversion of complex substrates to methane. With the exception of the hydrolysis
step, all other subprocesses of anaerobic treatment have been successfully modeled by
following Monod kinetics. The Contois and Chen & Hashimoto model has also been
used quite extensively to account for the effect of influent substrate concentration on
effluent quality. Based on a brief overview of the observed phenomena related to the
kinetics of mass transfer in methanogenesis, it is concluded that with but few
exceptions, the evidence for the significance of mass transfer effects in the different
reactor configurations is circumstantial and, in some cases, contradictory. Our
understanding of the kinetics of paniculate substrate removal in biofilms is still
incomplete for engineering applications, and more research is necessary.
Key words: Anaerobic digestion , Kinetics, Mass Transfer, Hydrolysis, Fermentation,
Anaerobic Oxidation, Methanogenesis, Acetogenesis, Waste water Sludge.

2.4 Thermophilic Anaerobic Digestion of Solid Waste for Fuel Gas Production
Chales L.Cooney ,Donald L. Wise
Anaerobic digestion offers a potential means of converting organic solid waste into
fuel gas and thereby provide a supplemental and readily utilizable source of energy.
4
We are particularly interested in the use of thermophilic digestion over a mesophilic
operation for it can achieve higher rates of digestion, greater conversion of waste
organics to gas, faster solid–liquid separation, and minimization of bacterial and viral
pathogen accumulation. Our results comparing mesophilic (37°C) and thermophilic
(65°C) anaerobic digestion of domestic solid waste confirm the increased rate and
conversion of waste to methane. In addition, utilizing radioactive labeling of glucose
and acetic acid, we have measured the volumetric rates of volatile acid production and
disappearance under both mesophilic and thermophilic conditions

CHAPTER-3
METHDOLOGY
3.1 Anaerobic digestion process

5
Anaerobic digestion is a collection of processes by which microorganisms break
down biodegradable material in the absence of oxygen. The process is used for
industrial or domestic purposes to manage waste or to produce fuels. Much of
the fermentation used industrially to produce food and drink products, as well as
home fermentation, uses anaerobic digestion. This happens in a tank called anaerobic
digester
Anaerobic digestion occurs naturally in some soils and in lake and oceanic
basin sediments, where it is usually referred to as "anaerobic activity". This is the
source of marsh gas methane as discovered by Alessandro Volta in 1776.
The digestion process begins with bacterial hydrolysis of the input materials.
Insoluble organic polymers, such as carbohydrates, are broken down to soluble
derivatives that become available for other bacteria. Acidogenic bacteria then convert
the sugars and amino acids into carbon dioxide, hydrogen, ammonia, and organic
acids. These bacteria convert these resulting organic acids into acetic acid, along with
additional ammonia, hydrogen, and carbon dioxide. Finally, methanogensconvert
these products to methane and carbon dioxide. The methanogenic archaea populations
play an indispensable role in anaerobic wastewater treatments.
Anaerobic digestion is used as part of the process to treat biodegradable waste
and sewage sludge. As part of an integrated waste management system, anaerobic
digestion reduces the emission of landfill gas into the atmosphere. Anaerobic
digesters can also be fed with purpose-grown energy crops, such as maize.
Anaerobic digestion is widely used as a source of renewable energy. The process
produces a biogas, consisting of methane, carbon dioxide, and traces of other
‘contaminant’ gases.This biogas can be used directly as fuel, in combined heat and
power gas engines] or upgraded to natural gas-quality biomethane. The nutrient-
rich digestate also produced can be used as fertilizer.
With the re-use of waste as a resource and new technological approaches that have
lowered capital costs, anaerobic digestion has in recent years received increased
attention among governments in a number of countries, among these the United
Kingdom (2011), Germany and Denmark (2011).

6
 Fig:3.1 Anaerobic Digester Model
3.2 Types of anaerobic digester:
The following is a partial list of types of anaerobic digesters. These processes and
systems harness anaerobic digestion for purposes such as sewage
treatment and biogas generation. Anaerobic digesters can be categorized according to
two main criteria: by whether the biomass is fixed to a surface ("attached growth") or
can mix freely with the reactor liquid ("suspended growth"); and by the organic
loading rate (the influent mass rate of chemical oxygen demand per unit volume). The
widely used UASB reactor, for example, is a suspended-growth high-rate digester,
with its biomass clumped into granules that will settle relatively easily and with
typical loading rates in the range 5-10 kgCOD/m3/d.

Examples of anaerobic digesters include:

 Anaerobic activated sludge process
 Anaerobic contact process
 Anaerobic expanded-bed reactor
 Anaerobic filter
 Anaerobic lagoon
 Batch system anaerobic digester
 Continuous stirred-tank reactor (CSTR)
 Expanded granular sludge bed digestion (EGSB)
 Internal circulation reactor (IC)
 One-stage anaerobic digester
 Plug-flow anaerobic digester

7
 Submerged media anaerobic reactor
 Two-stage anaerobic digester
 Upflow anaerobic sludge blanket digestion (UASB)
 Upflow and down-flow anaerobic attached growth
3.3 Working principle:
Biogas is produced through the anaerobic digestion (fermentation) of decaying
plant or animal matter. It is the naturally occurring emission of bacteria that
thrive without oxygen, and occurs in three steps. First is the decomposition, or
hydrolysis, of the biodegradable material into molecules such as sugars. Next,
these molecules are converted into acids. Lastly, the acids are converted into
biogas.

Fig: 3.3 Working Principle of Anaerobic Digester

3.4 Anaerobic Waste Treatment Processes

Advantage of anaerobic waste treatment systems as means for recovery of non-
conventional energy is increasingly being recognized worldwide. Anaerobic
decomposition is a biologically mediated process, indigenous to nature, and capable
of being simulated for treating wastes emanating from municipal, agricultural, and
industrial activities. Anaerobic digestion as applied in treatment of sewage sludge and
other organic wastes, represents the controlled application of a process. Although,
anaerobic digesters have traditionally been used for many decades in the stabilization
of sewage sludge, their successful and economic employment for the treatment of
liquid wastes is only a recent phenomenon, arising from the development of new
reactor designs. These concepts have led to development of various reactors, which

8
 are capable of retaining a much higher biomass concentration than traditional
digesters. Making the sludge retention independent of the influent retention time
makes this possible. The technological approaches to allow this condition of
independent sludge retention time can be divided in to the following:
 Attachment of biomass on the media (filters, fluidized systems, and RBC
configurations);
 Non-attached biomass concept as suspended growth process (sludge blanket reactors
and contact process with sludge recycling).
It is difficult to evaluate the advantages and disadvantages of each system in relation
to other concepts, as generalizations are not usually valuable in practice.
Considerations such as, purification rates, loading rates, investment cost, energy
balance, space requirements, operational costs, and specific long term experience with
certain wastewaters are all important, but they will be valued differently from
industryto industry. Various anaerobic reactor types in practice are summarized in
Table 3.4.1

Table-3.1 Basic Types of Reactors used in Anaerobic Process

9
Reference: Henze and Harremoes,1983.

In suspended growth systems microbial cultures are freely suspended in the reactors.
Microorganisms are suspended in the wastewater, as a single cell or as their
agglomerates. Various reactor types developed on the principles of suspended growth
are discussed hereunder. The anaerobic contact process essentially involves two
phases. A contact phase, where the raw waste is intimately mixed with a previously
developed and available anaerobic sludge culture in the reactor; and a separation
phase, where the active sludge particles are separated from the treated liquor and
recycled to the contact unit. In this process, raw wastewater is mixed with recycled
sludge solids and then digested in a digestion chamber. After digestion, the mixture is
separated in a clarifier (or vacuum flotation unit), and the supernatant is discharged as
effluent. Settled sludge is then recycled to seed the incoming waste.
The loading rates permissible in an anaerobic waste treatment process are primarily
dictated by the sludge retention in an anaerobic reactor. The maintenance of high
Sludge Retention Time (SRT) has been the major problem in the practical application
of the process, especially for waste with Chemical Oxygen Demand (COD) below
about 3000 mg/L. Obviously, a waste treatment process for low-strength wastes is an
economical one if large volume of waste can be forced through the system in a
relatively short time period. For this purpose process are required in which the
biomass retention time can be controlled independently of the wastewater flow rate.
Conventional anaerobic treatment processes of the flow-through type are therefore
inadequate to treat low-strength wastes.

10
Advances in the understanding of how anaerobic system function, improved
understanding of mixing and mass transfer, and anaerobic reactor design, has led to
the evolution of a new generation of high-rate anaerobic processes e.g. Anaerobic
Filters (AF), Anaerobic expanded / Fluidized bed rectors, Upflow Anaerobic Sludge
Blanket (UASB) Reactor, etc. These systems have been schematically presented in
Figure :3.2

Fig:3.2 Typical reactor configurations used in anaerobic wastewater treatment

One common feature offered by all the high-rate processes is their ability to provide
high SRT in relation to hydraulic retention time (HRT). High biomass concentration is
maintained in a reactor with relatively low treatment time. In anaerobic filter and
expanded / fluidized beds, this is accomplished by development of bio-film on support
surface. In UASB systems, this is accomplished by the development of granules or
flocs that have extremely good settling properties. Among the other improved high
rate anaerobic treatment methods, UASB has secured an important place.
Anaerobic Degradation of Organic matter
The factors that determine the removal efficiency of biodegradable organic matter are:
1. The nature and composition of the organic matter to be removed

11
2. Suitability of environmental factors
3. Sludge retention time in the reactor
4. The intensity of mixing, hence contact between bacterial biomass and organic
matter.
5. Specific loading of organic matter with respect to bacterial sludge mass, and
retention time.
Factors (1) and (2) are basically dependent on wastewater characteristics, whereas (3)
to (5) are related to the type and design of the treatment system. The transformation of
complex macromolecules of organic matter present in wastewater into biogas requires
several groups of microorganisms. The reaction sequence of the anaerobic digestion
of complex macromolecules is presented in Figure 3.4 [Gujer and Zehnder, 1983].
Different steps are necessary for the anaerobic digestion of proteins, carbohydrates,
and lipids. Four different phases can be distinguished in the overall conversion
process of organic matter to biogas as
1) Hydrolysis,
2) Acidogenesis,
3) Acetogenesis, and
4) Methanogenesis.

Fig:3.3 Stages of Anaerobic digestion.

12
Fig 3.4 Reaction Sequences for the Anaerobic Digestion of Complex Organic
Matter

3.5 Overview of Anaerobic Degradation Process

The anaerobic biological conversion of organic waste to methane is a complex
process involving a number of microbial populations linked by their individual
substrate and product specificities. The overall conversion process may be described
to involve direct and indirect symbiotic association between different groups of
bacteria. The product of one bacterium is often the substrate for others and hence, a
balance between the bacterial numbers and the substrate concentrations must be
maintained. The biological conversion of organic matter occurs in three steps. The
first step in the process involves transformation of highermolecular mass compounds
into compounds suitable for use as a source of energy and cell carbon (hydrolysis).
The second step (acidogenesis) involves the bacterial conversion of the compounds
resulting from the first step into identifiable lower-molecular-mass intermediate
compounds. Lower chain volatile fatty acids produced during acidogenesis are
utilized by a group of bacteria (acetogens) to produce acetate. The third step
(methanogenesis) involves the bacterial conversion of the intermediate compounds
into simpler end products, such as methane and carbon dioxide. Several
nomenclatures have been proposed for these three steps. Speece and McCarty (1962)
called the first and the second steps the constant BOD phase and the third step, the

13
reducing BOD phase, because only the methane formation in the third step brings
about the reduction of BOD or COD through the whole process.
According to trophic requirements the bacteria involved can be conveniently divided
into three groups as follows.
3.5.1 Hydrolytic bacteria - acidogens: These bacteria hydrolyze the substrate
(macromolecule) into short-chain organic acids and other small molecules, which can
be taken up and converted into soluble short-chain organic molecules, e.g.,
carbohydrates are converted into low-chain fatty acids, alcohols, hydrogen and carbon
dioxide under anaerobic condition.
Strict anaerobes are composed most part of this group of bacteria. The generation time
of these bacteria is 2 to 3 hours. The principle intermediate compounds resulting from
conversion of the substrate during acid fermentation are acetate (CH 3COOH),
propionate (CH3CH2COOH), butyrate (CH3CH2CH2COOH), hydrogen gas (H2),
carbon dioxide (CO2), lactate (CH3CHOHCOOH), formate (HCOOH), ethanol
(CH3CH2OH), valeric acid (CH3CH2CH2CH2COOH), isovaleric acid
((CH3)2CHCH2COOH), and caproic acid (CH3CH2CH2CH2CH2COOH). The
distribution of final product depends on the species of acidogenic bacteria and on the
environmental conditions such as pH and temperature.
3.5.2 Obligate Hydrogen Producing Acetogens (OHPA):
This group converts compound formed in the first stage into acetic acid and hydrogen.
Low hydrogen pressure favours these reactions [Harper and Pohland, 1986].
e.g. Propionate CH3CH2COOH + 2H2O → CH3COOH + CO2 + 3H2
ΔGo = 76.1 KJ/mole
Butyrate CH3CH2CH2COOH + 2H2O → 2CH3COOH + 2H2
ΔGo = 48.1 KJ/mole
From the viewpoint of the thermodynamics, a negative value of free energy change is
necessary for any reaction to proceed without input of external energy. This theory
apparently suggests that hydrogen producing acetogenic bacteria cannot obtain energy
for growth from these reactions. However, the value of free energy change in the
actual environment surrounding the bacteria, ΔG', is different from that of ΔGo' and
depends on the concentrations of substrates and products as follows [Harper and
Pohland,1986]:
[P1].[P2]....
ΔG' = ΔGo' + RTA ln ------------------ ....(1)

14
[S1].[S2]....
Where,
ΔG' = free energy change at pH = 7 (kJ/mol),
ΔGo' = standard free energy change at pH = 7 (kJ/mol),
R = gas constant = 0.082 L.atm/mol.oK,
TA = temperature (oK),
[P1].[P2]... = product concentration (mol/L or atm), and
[S1].[S2]... = substrate concentration (mol/L or atm).
Only low partial pressure of hydrogen can give negative values of ΔG' in above
equations, because substrate concentration cannot be so high and acetate
concentration is not so low in anaerobic reactors. This shows that extremely low
partial pressure of hydrogen is essential for hydrogen producing acetogenic bacteria
although they themselves produce hydrogen.
Experimentally it was found that the hydrogen partial pressure higher than 5 x 10-3
atm ceased the degradation of propionate by hydrogen producing acetogenic bacteria
[Hanaki et al., 1985]. Based on thermodynamics associated with this reactions Harper
and Pohland [1985] indicated that propionic acid oxidation to acetate becomes
favourable only at hydrogen partial pressure below 10-4 atm, while, butyric acid
oxidation becomes favourable at 10-3 atm H2 or below.
Hydrogen utilizing methanogenic bacteria can serve such a thermodynamically
favourable conditions for hydrogen producing acetogenic bacteria in anaerobic
reactors, thus, the activity of hydrogen producing acetogenic bacteria depends on the
existence of methanogenic bacteria. Hydrogen utilizing methanogens receive
hydrogen as a substrate from hydrogen producing acetogenic bacteria. The
interrelationship between these two groups of bacteria is called interspecies hydrogen
transfer, which also exists between acidogenic and methanogenic bacteria. Acidogenic
bacteria produce more hydrogen and acetate than propionate or lactate and obtain
more energy under low hydrogen partial pressure which is kept by methanogenic
bacteria. The interspecies hydrogen transfer is favourable but not essential for
acidogenic bacteria, while, it is indispensable for hydrogen producing acetogenic
bacteria.
3.5.3 Methanogenic bacteria - methanogens: These bacteria produce methane. The
doubling time of these bacteria is 2 - 10 days. These are further divided into two
groups as:

15
a) Hydrogen utilisers (lithotrophs)
CO2 + 4H2 →CH4 + 2H2O convert ADP to ATP
b) Acetic acid users (acetotrophs)
CH3COOH →CH4 + CO2 produce 0.25 mole of ATP
The methane producing bacteria are strict anaerobes which are extremely sensitive to
changes in temperature and pH. These bacteria are active in two temperature zones,
namely, in the mesophilic range (30oC - 35oC) and in the thermophilic range (50oC -
60oC). However, anaerobic processes have been operated at 15oC successfully when
sufficient residence time for these bacteria was provided.
The majority of methanogens in anaerobic wastewater treatment and natural anaerobic
environment utilize hydrogen and single carbon compounds as substrates for methane
production. In addition there are two known genera of methanogens which can utilize
the two-carbon compound, acetic acid. These include species of Methanosarcina and
Methanothrix (Methanosaeta). The Methanothrix species are unable to use hydrogen
in combination with CO2 and these are non-hydrogen-oxidizing acetotrophs (NHOA).
In contrast, Methanosarcina can utilize H2/CO2 as well as acetate, carbon monoxide,
methanol, and methylamines as growth substrates. Due to their ability to use both
H2/CO2 and acetate, these bacteria are classified as Hydrogen Oxidizing Acetotrophs
(HOA). Hydrogen- Oxidizing Methanogens (HOM) do not cleave acetate, but utilize
H2/CO2 and formate as substrates [Harper and Pohland, 1986]. The HOA are unique
in their capability to utilize multiple (one and/or two carbon) substrates. This ability
affords a higher potential for survival when competing with sulfur reducing bacteria
(SRB) and nitrate reducing bacteria (NRB) for hydrogen and acetate. At hydrogen
partial pressure >10-4 atm, HOA use H2/CO2 in favour of acetate, whereas acetate
cleavage by NHOA is unaffected by hydrogen. NHOA have a much higher affinity
for acetate than the HOA. NHOA may outcompete HOA at acetate concentrations
below 50 mg/L, while above 250 mg/L acetate, the HOA are more competitive
[Speece et al., 1983]. As a result of this comparative kinetics Methanothrix (NHOA)
may be found in reactors with lower organic loading. Methanosarcina are more
predominant in low retention time reactors such as in the lower reaches of plug flow
anaerobic filters and in two phase reactor system.
Oxidation of reduced organic products to bicarbonate and acetate also occurs due to
NRB and SRB. Higher organic waste conversion rates may be available through SRB
than through methanogenesis. Moreover, SRB and NRB are not limited to one-and

16
two-carbon substrates, as are methanogens. However, from process engineering
perspective, such an approach has disadvantages, including the loss of energy
available from methane and the production of hydrogen sulphide or ammonia. Since,
sulphide and ammonia are much more soluble than methane, their dissolved
components can contribute significantly to effluent COD [Harper and Pohland, 1986].
However, this approach may hold possibilities for reducing propionic acid and
hydrogen, as well as acetic acid in a stressed reactor, in order to more rapidly
reestablish the equilibrium with the existing hydrogen removal system.

17
 CHAPTER-4
FACTORS AFFECTING ANAEROBIC DIGESTION
Development of anaerobic process technology is dependent on a better understanding
of the factors that are associated with the stability of the biological processes
involved. Process instability is usually indicated by a rapid increase in the
concentration of volatile acids in the first stage of the reaction. Low pH with a
concurrent reduction in methane gas production indicates the methanogenesis more
susceptible to upset. Acclimatization of the microbes to a substrate may take 3 to 8
weeks. Sufficiently acclimated bacteria show greater stability towards stress-inducing
events such as hydraulic overloads, fluctuations in temperature, volatile acid and
ammonia concentrations, etc. Several environmental factors can affect anaerobic
digestion such as specific growth rate, decay rate, gas production, substrate utilization,
etc. The environmental factors of primary importance are discussed below.
4.1 pH, Acidity and Alkalinity
Methanogenic microorganisms are susceptible to the minute changes in the pH values.
Optimum pH range of 6.6 – 7.6 is considered favourable for the methane producing
bacteria, which cannot tolerate the fluctuations. The non-methanogenic bacteria do
not exhibit such strong sensitivity for environmental conditions and are able to
function in a range of pH form 5 – 8.5. The pH maintained inside the reactor, due to
the process results from the interaction of the carbon dioxide-bicarbonate buffering
system and volatile acids-ammonia formed by the process. It is necessary to prevent
the accumulation of acids to a level, which may become inhibitory to the
methanogenic bacteria. For this, it is important that there should be sufficient
buffering capacity present in the reactor, which may prevent the reactor from souring.
Although, the carbonates and bicarbonates of sodium and calcium are required to be
added to the digesters to provide buffering action, lime (Calcium hydroxide) is most
commonly used for this purpose. Only the unionized volatile acids in the
concentration range of 30 - 60 mg/L are toxic.
4.2 Temperature
As in all biological processes, anaerobic processes are affected by temperature. The
higher the temperature, higher is the microbial activity until an optimum temperature
is reached. A further increase of the temperature beyond its optimum value results in
steep decrease in activity. Anaerobic process can take place over a wide range of

18
temperatures (4 – 600C). Once as effective temperature is established, small
fluctuations can result in a process upset. Although most of the sludge digester are
operated in the mesophilic range (30 – 400C), methanogenesis can occur at
temperatures as low as 12 to 150C. The effect of increasing temperature on
biochemical reaction rate in the range of 4 – 250C is profound.
The optimum temperature for growth of anaerobic microorganisms is 350C or greater.
Although anaerobic digesters have been reported to operate at substantially lower
temperatures, such as 200C, anaerobic growth under these temperature conditions is
slow requiring prolonged start-up time and difficulties in operation. In situations
where reactor’s operating temperature is low, start-up will be benefited if initiated at
approximately 350C. At temperature of less than 25 0C, the digestion rate decreases
sharply and conventional anaerobic reactors in operation at ambient temperatures in
cooler climates may require detention times of as much as 12 weeks for the treatment
of sewage sludges.
The majority of industrial digester systems operate in the mesophilic range of 30 –
400C. It is probable that increase in microbial reaction rates at the elevated
temperatures of thermophilic processes (50o C– 600C), and hence decrease in SRT
may prove advantageous under some circumstances. However, lack of stability in
thermophilic municipal waste treatment can occur. Thermophilic digestion is most
practical where wastewater stream to be treated is discharged at an extremely high
temperature and the digester is present on site.
In psycrophilic, mesophilic, or thermophilic ranges, uniformity of temperature over
the entire vessel contents is of paramount importance to anaerobic digestion.
Temperature change of even a few degrees can result in a marked upset in microbial
metabolism and rapid alterations in reactions in the reactor and may necessitate
several days for the recovery. A consistent temperature throughout the system can be
provided by adequate mixing of the reactor by paddle, gas sparging, or flow over heat
exchangers.
4.3 Nutrients
Anaerobic wastewater treatment processes are often used for industrial waste with
only minor amount of nutrients present. This might result in nutrient deficiency,
unless additional nutrients are supplemented. Often the COD/N ratio and COD/N/P
ratio is used to described the nutrient requirements. Optimum N/P ratio can be
considered to be 7. The theoretical minimum COD/N –ratio is considered to be 350/7.

19
A value around 400/7 is considered reasonable for high rate anaerobic processes
(operated in SLR of 0.8 – 1.2 kg COD /kg VSS.d). For low rate processes (<0.5 kg
COD /kg VSS.d) the COD/N-ratio has been observed to be increased dramatically to
values of 1000/7 or more. Other than nitrogen and phosphorous, trace metals also are
essential for anaerobic processes.
The presence of trace metals such as molybdenum, selenium, tungsten and nickel is
probably necessary for the activity of several enzyme systems. When these trace
elements are not present in the wastewater, addition of nickel, cobalt, and
molybdenum can increase methane production and allow greater volumes of
wastewater to be effectively treated by decreasing the reactor residence time.
4.4 Inhibitory Substances
Inhibition of the anaerobic digestion process can be mediated to varying degrees by
toxic materials present in the system. These substances may be components of the
influent wastewater or by products of the metabolic activities in the digester.
Inhibitory toxic compounds include sulphides, consequential in the processing of
waste from sources such as molasses fermentation, petroleum refining and tanning
industries. Volatile acid and other microbial products can accumulate and inhibit
reactor-buffering capacity. Inhibition may also arise as the consequence of the
increased levels of ammonia, alkali, and the alkaline earth metals, and heavy metals in
the system.
4.5 Volatile Acids Inhibition:
Anaerobic reactor instability is generally evident by a marked and rapid increase in
volatile fatty acids concentrations; this is frequently indicative of the failure of the
methanogenic population due to other environmental disruptions such as shock
loadings, nutrient depletion or infiltration of inhibitory substances. Acetate has been
described as the least toxic of the volatile acids, while propionate has often been
implicated as a major effecter of digester failure. The inhibition by the volatile acids
at acidic pH values can be attributed to the existence of unionized VFAs in significant
quantities in the system. The undissociated nature of these acids allow them to
penetrate the bacterial cell membrane more efficiently than their ionized counterparts,
and once assimilated, induce an intracellular decrease in pH and hence a decrease in
microbial metabolic rate. The resulting VFA concentration in the reactor should be
maintained below 500 mg/L at any point of time and preferably below 200 mg/L for
optimum performance.

20
4.6 Ammonia – Nitrogen Inhibition: Although ammonia is an important buffer in
anaerobic processes, high ammonia concentration can be a major cause of operational
failure. In reactor system that has not previously been acclimated to high ammonia
loadings, shock loadings of high ammonia concentration generally caused rapid
production of VFAs such that the buffering capacity of the system may not be able to
compensate for the decrease in pH.
Further depression of alkalinity and reduction of pH may result in reactor failure.
Inhibition is indicated by a decrease in gas production and an increase in volatile acid
formation.
4.7 Sulphide Inhibition: The sulphate and other oxidized compounds of sulphur are
easily reduced to sulphide under the conditions prevalent in anaerobic digesters.
Sulphur-containing amino acids of protein can also undergo degradation to sulphide.
These compounds are of significance when anaerobic treatment is considered for
industrial processes which tend to produce large quantities of sulphides in their waste
stream. These sulphides formed by the activity of reactor microorganisms may be
soluble or insoluble, depending upon their associated cations. When the salts formed
are insoluble, they have negligible effects on digestion. Iron addition, for example can
suppress sulphide inhibition by removing S2- ion from solution by precipitation.
Desulfovibrio and other sulphate-reducing genera form sulphides from sulphates and
some of the fermentative microorganisms utilize the sulphur containing amino acids
to produce sulphides. Sulphide concentration in excess of 200 mg/L in a digester at 35
0C, with continuous feeding and mixing, produced severe inhibitory effects including
the complete cessation of gas production [Parkin and Speece, 1983]. All the heavy
metals, with the exception of chromium, form insoluble sulphide salts and thus can be
removed from solution by sulphide present in the system by precipitation. Free
sulphide can also be eliminated as hydrogen sulphide by vigorous gas production.
4.8 Heavy Metals Inhibition: The most common agents of inhibition and failure of
sewage sludge digesters are identified to be contaminating heavy metals. Heavy
metals in the soluble state are in general regarded to be of more significance to reactor
toxicity than are insoluble forms. Anaerobic digestion also reduces the valence states
of some heavy metals. Both copper and iron may be reduced from the trivalent to
divalent state. This reduces the quantity of the precipitating agent, such as sulphide,
necessary for the removal of the metal ion from solution. The heavy metals can be
removed from anaerobic systems by adsorption. Those digesters, such as the CSTR

21
configuration; which are tolerant to the wastes containing high levels of suspended
solids are effective in metal removal, provided sufficient adsorption sites are present.
The metals like copper, chromium, nickel, lead can induce toxicity in the reactor
when present in higher concentration, and acceptable concentration in the wastewater
to be treated differs from metal to metal.

Fig 4.1 Anaerobic digester plant

22
 CHAPTER-5
DESIGN OF ANAEROBIC DIGESTER
Anaerobic treatment of high solids such as animal manure, biological sludge,
nightsoil, etc. is commonly known as anaerobic digestion and is carried out in airtight
container known as anaerobic digester (AD).
• AD is usually continuous flow stirred tank reactor (CFSTR) for which HRT and
SRT is nearly the same i.e the ratio of SRT/HRT = 1.
• Design is based on volatile solids (VS) loading rate
Anaerobic treatment of wastewaters requires long SRT to achieve better treatment
efficiency
• The ratio of SRT/HRT ~ 10-100
• The high ratio allows the slow growing methanogens to remain in the reactor for
longer time

Fig: 5 .1 Design of anaerobic digester

Anaerobic Process Design

5.1 Design based on volumetric organic loading rate (VOLR):
VOLR = So.Q/V
VOLR : Volumetric organic loading rate
(kg COD/m3-day)
So : Wastewater biodegradable COD (mg/L)
Q : Wastewater flow rate (m3/day)
V : Bioreactor volume (m3)

23
So and Q can be measured easily and are known upfront VOLR can be selected!
How do we select VOLR?
 Conducting a pilot scale studies
 Find out removal efficiency at different VOLRs
 Select VOLR based on desired efficiency
5.2 Design based on hydraulic loading rate:

H : Reactor height (m)

. : Allowable hydraulic retention time (hr)

Q : Wastewater flow rate (m3/hr)
A : Surface area of the reactor (m2)
Permissible superficial velocity (Va)
Va=H/
For dilute wastewater with COD < 1000 mg/L
5.3 Design Factors
Anaerobic digester is designed in terms of size by using various approaches. Some
approaches are outlined below:
1. Solids retention time (SRT) : denoted by (days)
2. Volatile solids loading rate : kg VS/m3-day
3. Volume reduction
4. Loading factors based on population
Table:5.2 Important design parameters for anaerobic digesters

24
5.3.1 Solids (SLUDGE) retention time (SRT) in a CSTR
Completely stirred anaerobic reactor (CSTR) is a completely mixed reactor for which
solid retention time (SRT) and hydraulic retention time (HRT) is the same.
Influent flow rate (Q), m3/day
HRT, days =Volume V (m3)/ day
Flow rate Q (m3/day)
For a given SRT (HRT), the size of reactor can be easily determined since flow rate
(Q) is known to us.
Digester volume, V (m3) = Flow rate (Q) x SRT
Influent flow rate (Q), m3/day

Fig:5.3 Design based on Sludge Retension Time

5.3.2 Volatile solids loading rate
The size of an anaerobic digester can also be estimated based on volatile solids
loading rate expressed as kg VS/m3-day.
Volatile solids loading rate,= Influent VS (kg/day)/ Reactor volume (m3)
(kg VS/m3- day)
For a given volatile solids loading rate, the size of reactor can be easily determined
since influent VS (kg/day) is known to us.
Digester volume, V (m3) = Influent VS (kg/day)/Volatile solids loading rate,(kg
VS/m3- day)

Fig:5.4 Design Based on Volatile solids loading

25
Example:
How much methane gas can be generated through complete anaerobic
degradation of 1 kg COD at STP ?
Step 1: Calculation of COD equivalent of CH4
CH4 + 2O2 ------------------> CO2 + 2H2O
16 g 64g
=> 16 g CH4 ~ 64 g O2 (COD)
=> 1 g CH4 ~ 64/16 = 4 g COD ------------ (1)

Step 2: Conversion of CH4 mass to equivalent volume

Based on gas law, 1 mole of any gas at STP (Standard Temperature and Pressure)
occupies volume of 22.4 L.
=> 1 Mole CH4 ~ 22.4 L CH4
=> 16 g CH4 ~ 22.4 L CH4
=> 1 g CH4 ~ 22.4/16 = 1.4 L CH4 ---------- (2)
Step 3: CH4 generation rate per unit of COD removed
From eq. (1) and eq. (2), we have,
=> 1 g CH4 ~ 4 g COD ~ 1.4 L CH4
=> 4 g COD ~ 1.4 L CH4
=> 1 g COD ~ 1.4/4 = 0.35 L CH4
or 1 Kg COD ~ 0.35 m3 CH4 ----------- (3)
complete anaerobic degradation of 1 Kg COD produces 0.35 m3 CH4 at STP .

26
 CHAPTER-6
ADVANTAGES, DISADVANTAGES, LIMITATIONS
AND APPLICATIONS
6.1 Advantages:

 It is a net energy producing process which produces renewable energy in the

form of biogas.

 It produces a liquid and a fibrous fertiliser.

 It reduces odour below unprocessed waste odour levels.

 It is much less likely to cause environmental pollution than spreading

untreated organic waste on land.

 Reduction in number of pathogens.

 Low operating costs.

 Replacement of Fossil fuels

 Reducing vehicle movements

 Reducing usage of LP Gas for cooking

 Reducing or eliminating the energy footprint of waste treatment plants

 Reducing Methane emission from landfills

 Reducing electrical grid transportation losses

 Displacing industrially produced chemical fertilizers

 An important component of the zero waste initiatives

6.2 Disadvantages:

 When carried out at a commercial scale on farms and at wastewater treatment

works it requires a high level of investment in large tanks and other process
vessels.

 If run inefficiently AD can cause an odour nuisance.

 Not easily controllable

 Needs constant supervision.

 Maintenance and cleaning is difficult.

27
  Formation of explosive gases.

 It is a slow process.

6.3 Limitations:

1. Long start-up time

Because of lower biomass synthesis rate, it requires longer start-up time to attain a
biomass concentration.
2. Long recovery time
If an anaerobic system subjected to disturbances either due to biomass wash-out, toxic
substances or shock loading, it may take longer time for the system to return to
normal operating condition.
3. Specific nutrients/trace metal requirements
Anaerobic microorganisms especially methanogens have specific nutrients
e.g. Fe, Ni, and Co requirement for optimum growth.
4. More susceptible to changes in environmental conditions
Anaerobic microorganisms especially methanogens are prone to changes in conditions
such as temperature, pH, redox potential,etc.
6.4 Applications:
 Domestic
 Industrial
 Waste and waste water treatment
 Electricity generation
 Grid generation
 Fertiliser and soil conditioner

28
 CHAPTER-7

OPERATION AND MAINTENANCE

Operation of an anaerobic digester will require more maintenance than other manure
management practices, such as composting or waste lagoon management. Installation
of an anaerobic digester may require hiring 1 to 2 additional employees for routine
maintenance, depending on the size of the operation. Be prepared to meet additional
maintenance requirements if you are considering anaerobic digester installation. Some
of the common maintenance activities are listed below with the frequency requirement
in parenthesis.
• Sludge Removal (every 1-2 years) – An anaerobic digester system must be cleaned
and removed of excess sludge. In well-designed systems, this is performed
automatically with very little downtime. Other designs require manual removal of
waste.
• Pump Clearing (every 3-6 months) – When pumping high solids content waste, it is
important to ensure that pumps are cleared of debris regularly. Items such as cow tails
(when removed for ease of milking), sand, work tools and other inorganic substances
can clog pumps hindering operation of the digester.
• Iron Packing Replacement (every 6-12 months) – It is important to remove the
corrosive hydrogen sulfide compounds to avoid engine replacement if biogas
collected from the digesters is being refined and used for electricity generation. This
can be done by passing the biogas through iron packing material. The iron packing
should be replaced at least every 12 months.
• General Engine Maintenance (every week) – Just as in your car, the generator
producing electricity from the anaerobic digester must be inspected for proper fluid
levels.
• Preventative Engine Maintenance (every month) – The electrical, fuel and air intake
systems must also be inspected for each of the gen sets.
• Valve Leak Checks (every 6-12 months) – To avoid safety hazards, it is
recommended that the valves on the digestion system be checked for leaks one to two
times a year. Improperly working valves should be replaced as soon as possible.

29
 • Pipe Leak Checks (every 6-12 months) – Pipes must be checked for leaks at least
once per year. It is also important that no open flames are anywhere near inflow or
outflow pipe lines.
• Fittings Leak Checks (every 6-12 months) – Any non metal fitting (i.e. ducted vents,
plastic valves, rubber fittings) located on the gas or waste pipeline must be inspected.
Other maintenance activities may be required specific to the system in place. Make
sure to discuss maintenance requirements with your technology provider to ensure
that an adequate maintenance plan is put in place. Proper maintenance of your
anaerobic digester and related components will both extend the lifetime of the system
as well as save money over the long term. Successful anaerobic digester operation
depends on routine maintenance activities.

Table: 7.1 Anaerobic treatment plants in Sub-tropical regions installed by four

international companies
Country Number COD removal Typical application
Ton-COD/d
India 75 3377.9 Distillary, Tannery
Brazil 77 1046.2 Brewery
China 43 366.5 Beverage, Brewery,
Food
Mexico 22 413.4 Beverage, Coffee,
Brewery,
Food
Phillipine 22 454.4 Food, Brewery,
Distillery, Beverage
Taiwan 21 207.5 Food, Leachate,
Chemical beverage,
Brewery.
Thailand 14 552.5 Alcohol,Agro
industrial waste water
Indonesia 10 31.8
Colombia 9 146.3
Isreal 9 67.8
Venezuela 9 264.1
Malaysia 8 142.2
Chile, South Africa 3 each
Argentina, Vietnam 2 each
Hong kong, Saudi Arabia, 1 each
Pakisthan
Total 338 7476.1

30
 CHAPTER-8
CONCLUSIONS
Quantity of biological solids produced in the anaerobic systems per unit weight of
organic material is much less than that in aerobic systems. This is a major advantage
of the anaerobic process as the quantity of sludge for ultimate disposal is reduced.
This is a result of conversion of volatile solids present, to the high energy level end
products such as methane, carbon dioxide and water. Methane has a definite economic
value as a fuel, and it is used as a source of energy for both heat and power in many
installations.
Another major advantage is the loading potential. Aerobic processes are restricted in
maximum organic loading rate by the inability to transfer oxygen at the rate sufficient
to satisfy the oxygen demand of the systems. Such limitation in organic loading rate is
not there for anaerobic processes. The stabilized sludge from anaerobic process may
be free from strong or foul odours and can be used for land application as ultimate
disposal because the digested sludge contains sufficient nutrients required for plants.
Pathogens are also destroyed to a high degree during the thermophilic anaerobic
process. Due to large retention time and consequent low growth rate, the cell yield is
also extremely low; thus, most of the carbon in the waste is available for
methanogenesis and under normal circumstances the yield of methane would, on an
average, be 0.33 – 0.36 m3 per kg COD utilized at 35oC and atmospheric pressure.
However, anaerobic treatment processes are not largely being implemented, because
of many factors. Anaerobic microorganisms, especially methanogens have slow
growth rate. At lower HRTs, the possibility of washout of biomass is more prominent
due to higher upflow velocity. This makes it difficult to maintain the effective number
of useful microorganisms in the system. To maintain the population of anaerobes,
large reactor volume or higher HRTs with low upflow velocity is required. This may
ultimately provide longer SRTs more than 40 days for high rate systems. Thus,
provision of larger reactor volume or higher HRTs ultimately leads to higher capital
cost. Special attention is therefore required towards controlling the factors that affect
process adversely; importantly among them being environmental factors such
as,temperature, pH, and concentration of toxic substances. Hence, skill supervision is
required for operating anaerobic reactor at optimal performance.

31
 REFERENCES

1. A. C. Haandel and G. Lettinga, Anaerobic Sewage Treatment: A Practical

Guide for Regions with A Hot Climate, John Wiley and Sons, Chichester, UK, 3rd
edition, 1994. View at Publisher · View at Google Scholar
2. A. Farghaly and A. Tawfik, “Simultaneous hydrogen and methane production
through multi-phase anaerobic digestion of paperboard mill wastewater under
different operating conditions,” Applied Biochemistry and Biotechnology, vol. 181,
no. 1, pp. 142–156, 2017. View at Publisher · View at Google Scholar · View at
Scopus
3. E. Metcalf and I. Eddy, Wastewater Engineering Treatment and Reuse,
McGraw-Hill Education, New York, NY, USA, 4th edition, 2003.
4. G. Lettinga, S. Rebac, and G. Zeeman, “Challenge of psychrophilic anaerobic
wastewater treatment,” Trends in Biotechnology, vol. 19, no. 9, pp. 363–370,
2001. View at Publisher · View at Google Scholar ·View at Scopus
5. K. Yetilmezsoy and S. Sakar, “Development of empirical models for
performance evaluation of UASB reactors treating poultry manure wastewater under
different operational conditions,” Journal of Hazardous Materials, vol. 153, no. 1-2,
pp. 532–543, 2008. View at Publisher · View at Google Scholar ·View at Scopus
L. Seghezzo, Anaerobic Treatment of Domestic Wastewater in Subtropical Regions,
Wageningen University, Wageningen, The Netherlands, 2004.
6. L. Singh, Z. A. Wahid, M. F. Siddiqui, A. Ahmad, M. H. A. Rahim, and M.
Sakinah, “Application of immobilized upflow anaerobic sludge blanket reactor using
Clostridium LS2 for enhanced biohydrogen production and treatment efficiency of
palm oil mill effluent,” International Journal of Hydrogen Energy, vol. 38, no. 5, pp.
2221–2229, 2013. View at Publisher · View at Google Scholar · View at Scopus
7. M. Henze and A. Ledin, “Types, characteristics and quantities of classic,
combined domestic wastewaters,” 2004.
8. N. Mahmoud, Anaerobic pre treatment of sewage under low temperature
(15°C) conditions in an integrated UASB-Digester system, Department of
Environmental Technology and the Department of Agro Technology and Food
Sciences Wageningen University, Wageningen, The Netherlands, 2002.

32
9. P. S. James and S. Kamaraj, “Immobilized cell anaerobic bioreactors for
energy production from agro-industrial waste waters-An introduction,” Bioenergy
News, vol. 6, no. 3, article 10, 2002. View at Google Scholar

33