Principles of anaerobic digestion 2017

Contents
1. Principles of anaerobic digestion ................................................................................... 1
1.1 Overview of Anaerobic Degradation Process .............................................................. 1
2. Microbiology of anaerobic digestion .............................................................................. 2
a. Hydrolysis and acidogenesis ........................................................................................... 5
(b)Acetogenesis ................................................................................................................. 6
(c)Methanogens ................................................................................................................. 7
(d) Sulfate reduction ......................................................................................................... 8
3. Factors Affecting Anaerobic Digestion .......................................................................... 9
Temperature .................................................................................................................... 10
4.Merits of Anaerobic Decomposition Process ................................................................... 11
4. Anaerobic sludge digesters ........................................................................................... 13
References .......................................................................................................................... 16

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1. Principles of anaerobic digestion

1.1 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 higher molecular 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 (methanogens) 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 reducing BOD phase, because only the methane
formation in the third step brings about the reduction of BOD or COD through the whole process.
Anaerobic biodegradation of organic material proceeds in the absence of oxygen and the presence
of anaerobic microorganisms. AD is the consequence of a series of metabolic interactions among
various groups of microorganisms. It occurs in three stages, hydrolysis/liquefaction, acidogenesis
and methanogens. The first group of microorganism secretes enzymes, which hydrolyses
polymeric materials to monomers such as glucose and amino acids. These are subsequently
converted by second group i.e. acetogenic bacteria to higher volatile fatty acids, H2 and acetic acid.
Finally, the third group of bacteria, methanogenic, convert H2, CO2, and acetate, to CH4. These
stages are described in detail below.
Inorganic electron acceptors, such as SO4 2− or CO2, are used in the oxidation process of organic
matter under anaerobic conditions. Methane formation does not occur in mediums where oxygen,
nitrate or sulfate is readily available as electron acceptors. Methane production occurs in different
natural environments, such as swamps, soil, river sediments, lakes and seas, as well as in the

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digestive organs of ruminant animals, where the redox potential is around −300 mV It is estimated.
That anaerobic digestion with methane formation is responsible for the complete mineralization of
5 to 10% of all the organic matter available on the Earth.

Anaerobic digestion represents an accurately balanced ecological system, where different

populations of microorganisms present specialized functions, and the breakdown of organic
compounds is usually considered a two-stage process. In the first stage, a group of facultative and
anaerobic bacteria converts (by hydrolysis and fermentation) the complex organic compounds
(carbohydrates, proteins and lipids) into simpler organic materials, mainly volatile fatty acids
(VFA), as well as carbon dioxide and hydrogen gases.

In the second stage, the organic acids and hydrogen are converted into methane and carbon
dioxide. This conversion is performed by a special group of microorganisms, named methanogens,
which are strictly anaerobic prokaryotes.

The methanogenic archaea depend on the substrate provided by the acid-forming microorganisms,
consisting, therefore, in a syntrophic interaction.

The methanogens carry out two primordial functions in the anaerobic ecosystems: they produce
an insoluble gas (methane) which enables the removal of organic carbon from the environment,
and they also keep the H2 partial pressure low enough to allow conditions in the medium for
fermenting and acid-producing bacteria to produce more oxidized soluble products, such as acetic
acid. Once the methanogens occupy the terminal position in the anaerobic environment during
organic compound degradation, their inherent low growth rates usually represent a limiting factor
in the digestion process as a whole.

2. Microbiology of anaerobic digestion

Anaerobic digestion can be considered an ecosystem where several groups of microorganisms
work interactively in the conversion of complex organic matter into final products, such as methane,
carbon dioxide, hydrogen sulfide, water and ammonia, besides new bacterial cells.

Although anaerobic digestion is generally considered a two-phase process, it can be subdivided

into various metabolic pathways, with the participation of several microbial groups, each with a
different physiological behavior, as illustrated in Figure 1 and described in the following items.

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Figure 1. Metabolic pathways and microbial groups involved in anaerobic digestion

Adapted from: Lettinga et al. (1996)

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Steps in anaerobic digestion involving four groups of bacterial activities

A. Hydrolysis

B. Fermentation

C. Acetogenesis

D. Methanogenesis

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a. Hydrolysis and acidogenesis
In the first stage of hydrolysis, or liquefaction, fermentative bacteria convert the insoluble complex
organic matter, such as cellulose, into soluble molecules such as sugars, amino acids and fatty
acids. The complex polymeric matter is hydrolyzed to monomer, e.g., cellulose to sugars or
alcohols and proteins to peptides or amino acids, by hydrolytic enzymes, (lipases, proteases,
cellulases, amylases, etc.) secreted by microbes. The hydrolytic activity is of significant
importance in high organic waste and may become rate limiting. Some industrial operations
overcome this limitation by the use of chemical reagents to enhance hydrolysis. The application
of chemicals to enhance the first step has been found to result in a shorter digestion time and
provide a higher methane yield (RISE-AT, 1998).
Hydrolysis/Liquefaction reactions
Lipids → Fatty Acids
Polysaccharides → Monosaccharides
Protein → Amino Acids
Nucleic Acids → Purines & Pyrimidines
Since the microorganisms are not capable of assimilating particulate organic matter, the first phase
in the anaerobic digestion process consists in the hydrolysis of complex particulate material
(polymers) into simpler dissolved materials (smaller molecules), which can penetrate through the
cell membranes of the fermentative bacteria. Particulate materials are converted into dissolved
materials by the action of exoenzymes excreted by the hydrolytic fermentative bacteria. The
hydrolysis of polymers usually occurs slowly in anaerobic conditions, and several factors may
affect the degree and rate at which the substrate is hydrolyzed (Lettinga et al., 1996):
 operational temperature of the reactor

 residence time of the substrate in the reactor

 substrate composition (e.g. lignin, carbohydrate, protein and fat contents)

 size of particles

 pH of the medium

 concentration of NH4 + −N

 concentration of products from hydrolysis (e.g. volatile fatty acids)

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The soluble products from the hydrolysis phase are metabolized inside the cells of the fermentative
bacteria and are converted into several simpler compounds, which are then excreted by the cells.
The compounds produced include volatile fatty acids, alcohols, lactic acid, carbon dioxide,
hydrogen, ammonia and hydrogen sulfide, besides new bacterial cells.

Acidogenesis is carried out by a large and diverse group of fermentative bacteria. Usual species
belong to the clostridia group, which comprises anaerobic species that form spores, able to survive
in very adverse environments, and the family Bacteroidaceaea, organisms commonly found in
digestive tracts, participating in the degradation of sugars and amino acids.

(b)Acetogenesis
In the second stage, acetogenic bacteria, also known as acid formers, convert the products of the
first phase to simple organic acids, carbon dioxide and hydrogen.

The principal acids produced are acetic acid (CH3COOH), propionic acid (CH3CH2COOH),
butyric acid (CH3CH2CH2COOH), and ethanol (C2H5OH). The products formed during
acetogenesis are due to a number of different microbes, e.g., syntrophobacter wolinii, a propionate
decomposer and sytrophomonos wolfei, a butyrate decomposer. Other acid formers are clostridium
spp., peptococcus anerobus, lactobacillus, and actinomyces (www.biogasworks.com- Microbes in
AD).

An acetogenesis reaction is shown below: C6H12O6 → 2C2H5OH + 2CO2

Acetogenic bacteria are responsible for the oxidation of the products generated in the acidogenic
phase into a substrate appropriate for the methanogenic microorganisms. In this way, acetogenic
bacteria are part of an intermediate metabolic group that produces substrate for methanogenic
microorganisms. The products generated by acetogenic bacteria are acetic acid, hydrogen and
carbon dioxide.

During the formation of acetic and propionic acids, a large amount of hydrogen is formed,
causing the pH in the aqueous medium to decrease. However, there are two ways by which
hydrogen is consumed in the medium: (i) through the methanogenic microorganisms, that use
hydrogen and carbon dioxide to produce methane; and (ii) through the formation of organic acids,
such as propionic and butyric acids, which are formed through the reaction among hydrogen,
carbon dioxide and acetic acid.
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Among all the products metabolized by the acidogenic bacteria, only hydrogen and acetate can
be directly used by the methanogenic microorganisms. However, at least 50% of the biodegradable
COD are converted into propionic and butyric acids, which are later decomposed into acetic acid
and hydrogen by the action of the acetogenic bacteria.

(c)Methanogens
Finally, in the third stage methane is produced by bacteria called methane formers (also known as
methanogens) in two ways: either by means of cleavage of acetic acid molecules to generate carbon
dioxide and methane, or by reduction of carbon dioxide with hydrogen. Methane production is
higher from reduction of carbon dioxide but limited hydrogen concentration in digesters results in
that the acetate reaction is the primary producer of methane (Omstead et al, 1980).
The final phase in the overall anaerobic degradation process of organic compounds into methane
and carbon dioxide is performed by the methanogenic archaea. They use only a limited number of
substrates, comprising acetic acid, hydrogen/carbon dioxide, formic acid, methanol, methylamines
and carbon monoxide. In view of their affinity for substrate and extent of methane production,
methanogenic microorganisms are divided into two main groups, one that forms methane from
acetic acid or methanol, and the other that produces methane from hydrogen and carbon dioxide,
as follows:

 Acetate-using microorganisms (aceticlastic methanogens)

 Hydrogen-using microorganisms (hydrogen trophic methanogens)
Aceticlastic methanogens. Although only a few of the methanogenic species are capable of
forming methane from acetate, these are usually the microorganisms prevailing in anaerobic
digestion. They are responsible for about 60 to 70% of all the methane production, starting from
the methyl group of the acetic acid.

Two genera utilize acetate to produce methane: Methanosarcina prevails above 10−3 M acetate,
while Methanosaeta prevails below this acetate level (Zinder, 1993). Methanosaeta may have
lower yields and be more pH-sensitive, as compared to Methanosarcina (Schimidt and Ahring,
1996). Methanosarcina has a greater growth rate, while Methanosaeta needs a longer solids
retention time, but can operate at lower acetate concentrations. The Methanosaeta genus is
characterized by exclusive use of acetate, and having a higher affinity with it than the
methanosarcinas. They are developed in the form of filaments, being largely important in the

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formation of the bacterial texture present in the granules. The organisms belonging to the
Methanosarcina genus are developed in the form of coccus, which group together forming
“packages”. They are considered the most versatile among the methanogenic microorganisms,
since they own species capable of using also hydrogen and methylamines (Soubes, 1994).

Hydrogen trophic methanogens. Unlike the aceticlastic organisms, practically all the well-known
methanogenic species are capable of producing methane from hydrogen and carbon dioxide. The
genera more frequently isolated in anaerobic reactors are Methanobacterium, Methanospirillum
and Methanobrevibacter. Both the aceticlastic and the hydrogen trophic methanogenic
microorganisms are very important in the maintenance of the course of anaerobic digestion, since
They are responsible for the essential function of consuming the hydrogen produced in the
previous phases. Consequently, the partial pressure of hydrogen in the medium is lowered, thus
enabling the production reactions of the acidogenic and acetogenic bacteria.

(d) Sulfate reduction

In reactors treating wastewater containing sulfate or sulfite, these compounds can be used by
sulfate-reducing bacteria (SRB) as acceptors of electrons released during the oxidation of organic
materials (Lettinga et al., 1996).

The metabolism of SRB is important in the anaerobic process, mostly because of their end
product, hydrogen sulfide. SRB group species have in common the dissimilatory sulfate
metabolism under strict an aerobiosis, and are considered a very versatile group of microorganisms,
capable of using a wide range of substrate, including the whole chain of volatile fatty acids, several
aromatic acids, hydrogen, methanol, ethanol, glycerol, sugars, amino acids and several phenol
compounds. Two major metabolic groups of SRB can be distinguished: (i) a group of species that
is able to oxidize incompletely its substrates to acetate, like the genera Desulfobulbus sp. and
Desulfomonas sp., and most of the species of the genera Desulfotomaculum and Desulfovibrio
belong to this group; and (ii) a group which is able to oxidize its organic substrates, including
acetate, to carbon dioxide.

The genera Desulfobacter, Desulfococcus, Desulfosarcina, Desulfobacterium and Desulfonema

belong to this group.

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In the absence of sulfate, the anaerobic digestion process occurs according to the metabolic
sequences presented in Figure 1. With the presence of sulfate in the wastewater, many of the
intermediate compounds formed by means of the metabolic routes identified in Figure 1 start to be
used by the SRB, causing a change in the metabolic routes in the anaerobic digester. Hence, the
SRB start to compete with the fermentative, acetogenic and methanogenic microorganisms for the
substrate available, resulting in a decrease in the production of methane from a given amount or
organic material present in the influent. The importance of this bacterial competition is greater
when the relative concentration of SO4 2− is increased in relation to the COD concentration.

3. 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
methanogens 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.

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 favorable 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

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hydroxide) is most commonly used for this purpose. Only the unionized volatile acids in the
concentration range of 30 - 60 mg/L are toxic.
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 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),
methanogens 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 250C, 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 thermophiles
processes (50 – 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 psychrophilic, 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.

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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. 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 [Van den Berg and Lentz, 1980].
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.Merits of Anaerobic Decomposition Process

It has been recognized that the anaerobic treatment is in many ways ideal for wastewater treatment
and has several merits mentioned as below:
 A high degree of waste stabilization;
 A low production of excess biological sludge and this sludge can be directly dried on sludge
drying bed without further treatment due to better dewatering ability;
 Low nutrient requirements, hence anaerobic treatment is attractive for the treatment of
wastewater where external nutrient addition is required;
 No oxygen requirement, hence saving in power required for supply of oxygen in aerobic
methods;
 Production of valuable byproduct, methane gas;
 Organic loading on the system is not limited to oxygen supply hence higher loading rate as
compared to aerobic processes can be applied.
 Less land required as compared to many aerobic process.

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 Non-feed conditions for few months do not affect adversely to the system and this makes it
attractive option for seasonal industrial wastewater treatment.
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 methanogens 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 up flow 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 up flow
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. Low synthesis / reaction rate hence, long start-up periods and difficulty in recovery from
upset conditions are some of the notable disadvantages. 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.

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4. Anaerobic sludge digesters

Conventional digesters are mainly used for the stabilization of primary and secondary sludge,
originating from sewage treatment, and for the treatment of industrial effluents with a high
concentration of suspended solids. They usually consist of covered circular or egg-shaped tanks of
reinforced concrete. The bottom walls are usually inclined, so as to favor the sedimentation and
removal of the most concentrated solids. The covering of the reactor can be fixed or floating.
Since conventional digesters are preferably used for the stabilization of wastes with a high
concentration of particulate material, the hydrolysis of these solids can become the limiting stage
of the anaerobic digestion process. The hydrolysis rate, in turn, is affected by several factors, such
as: (i) temperature; (ii) residence time;
(iii) Substrate composition and (IV) particle size.
Thus, with the aim to optimize the hydrolysis of the particulate material, conventional digesters
may be heated up, with operation temperatures usually ranging from 25 to 35 ◦ C. The hydrolysis
phase evolves very slowly when the digesters are operated at temperatures below 20 ◦ C.
As the conventional digesters do not have specific means for biomass retention in the system,
the hydraulic detention time should be long enough to guarantee the permanence and
multiplication of the microorganisms in the system, while enabling all the phases of the anaerobic
digestion to be processed appropriately.
Depending on the existence of mixing devices and on the number of stages, three main digester
configurations have been applied:
 Low-rate anaerobic sludge digester
 One-stage high-rate anaerobic sludge digester
 Two-stage high-rate anaerobic sludge digester
(a) Low-rate anaerobic sludge digester
The low-rate digester does not have mixing devices and usually comprises a single tank, where the
digestion, sludge thickening and supernatant formation occur simultaneously. Raw sludge is added
to the part of the digester where the sludge is undergoing active digestion and the biogas is being
released. With the up flow movement of the biogas, particles of sludge and other flotation materials
are taken to the surface, forming a scum layer. As a result of the digestion, the sludge stratifies
below the scum layer, and four different zones are formed inside the reactor, as characterized:
scum zone, supernatant zone, active digestion zone and stabilized sludge zone.

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The supernatant and stabilized sludge are periodically removed from the digester. Because of the
sludge stratification and the absence of mixing, no more than 50% of the digester volume are
actually used in the digestion process, with large reactor volumes being required to achieve good
sludge stabilization. In view of these limitations, low-rate digesters are mainly used in small
treatment plants.

Figure 1. Schematic representation of a low-rate anaerobic sludge digester

b) One-stage high-rate anaerobic sludge digester

The one-stage high-rate digester incorporates supplemental heating and mixing mechanisms,
besides being operated at uniform feeding rates and with the previous thickening of the raw sludge,
to guarantee more uniform conditions in the whole digester. As a result, the tank volume can be
reduced and the stability of the process is improved. Figure 4.2 presents a schematic representation
of a one-stage high-rate digester.
The solids retention times recommended for the design of complete-mix di- gesters are
illustrated in Figure below, and the high dependence of these in relation to the operational
temperature of the digester can be observed. When sizing the reactor, the hydraulic detention time
shall be equal to the solids retention time, as the system does not have a solids retention mechanism.

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Figure 4.2. Schematic representation of a one-stage high-rate anaerobic sludge digester

(C) Two-stage high-rate anaerobic sludge digester

Basically, the two-stage digester consists in the incorporation of a second tank, operating in series
with a high-rate primary digester, as illustrated in Figure 4.4.
In this configuration, the first tank is used for the digestion of the sludge, and may therefore be
equipped with heating and mixing devices. The second tank is used

Figure 4.4. Schematic representation of a two-stage high-rate anaerobic sludge digester

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References
Gavala, H.N., Yenal, U., Skiadas, I.V., Westermann, P. and Ahring, B.K. (2003) Mesophilic and
Thermophilic Anaerobic Digestion of Primary and Secondary Sludge, Effect of Pretreatment at
Elevated Temperatures. Water Research, 37, 4561-4572.
Rozzi, A. and Remigi, E. (2004) Methods of Assessing Microbial Activity and Inhibition under
Anaerobic Conditions: A Literature Review. Re/Views in Environmental Science &
Bio/Technology, 3, 93-115.
http://dx.doi.org/10.1007/s11157-004-5762-z
Problems-related operations-based education (probe), water Environment federation, alexandria,
va Toni glymph, author. American water works association, denver, co.
Wpcf manual of practice fd-13/asce manuals and reports on engineering practice no. 63.
Author bill marten, triad engineering inc. Series of six articles in wisconsin wastewater
Operator association's the clarifier, volumes 159-164.
Www.wef.org
Http://www.owp.csus.edu/ or available through inter-library loan at
Http://aqua.wisc.edu/waterlibrary
Www.awwa.org
Enve 302 environmental engineering unit processes by assist. Prof. Bilge alpaslan kocamemi
Marmara university department of environmental engineering istanbul, turkey
Rick snyder, governor dan wyant, director www.michigan.gov/deq 800-662-9278 environmental
assistance center

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