TYPES OF BACTERIA ON THE BASIS OF PROCESS MICROBIOLOGY

Fermentative bacteria: This group of bacteria is responsible for the first stage of anaerobic
digestion - hydrolysis and acidogenesis. Fermentation bacteria are anaerobic, but use organic
molecules as their final electron acceptor to produce fermentation end-products. Streptococcus,
Lactobacillus, and Bacillus, for example, produce lactic acid, while Escherichia and Salmonella
produce ethanol, lactic acid, succinic acid, acetic acid, CO2, and H2. Fermenting bacteria have
characteristic sugar fermentation patterns, i.e., they can metabolize some sugars but not others.
For example, Neisseria meningitidis ferments glucose and maltose, but not sucrose and lactose,
while Neisseria gonorrhoea ferments glucose, but not maltose, sucrose or lactose. Such
fermentation patterns can be used to identify and classify bacteria. The anaerobic species
belonging to the family of Streptococcaceae and Enterobacteriaceae and to the genera of
Bacteroides, Clostridium, Butyrivibrio, Eubacterium, Bifidobacterium and Lactobacillus are
most common.
Hydrogen producing acetogenic bacteria: Acetogenic bacteria are a specialized group of
strictly anaerobic bacteria that are ubiquitous in nature. Together with the methane‐forming
archaea they constitute the last limbs in the anaerobic food web that leads to the production of
methane from polymers in theabsence of oxygen. Acetogens are characterized by a unique
pathway, the Wood– Ljungdahl pathway of carbon dioxide reduction with the acetyl‐CoA
synthase as the key enzyme. This pathway also allows chemolitho-autotrophic growth on
hydrogen and carbon dioxide and it is the only pathway known that combines carbon dioxide
fixation with adenosine triphosphate (ATP) synthesis. Thus, it is considered the first biochemical
pathway on earth. ATP is synthesized by a chemi-osmotic mechanism with Na+ or H+ as
coupling ion, depending on the organism. In cytochrome‐free acetogens, energy is conserved by
reduction followed by dependent Na+ (or H+ ) translocation across the membrane (Rnf
complex). Acetogens may represent ancestors of the first bio energetically active cells in
evolution.
Homoacetogenes: The homoacetogens are much more adaptable than methanogens because in
addition to being autotrophic they can also live as chemoheterotrophs. Clostridium aceticum and
Acetobacterium woodii are the two homoacetogenic bacteria isolated from the sludge. In the
heterotrophic growth mode they can ferment glucose and derive some ATP by substrate level
phosphorylation. In so doing they generate carbon dioxide and hydrogen which can then be used
to power the chemiosmotic mechanism which allows them to derive some ATP also by anaerobic
respiration. The overall stoichiometry of this growth mode of homoacetogens is shown below.
Methanogens: Methanogens are autotrophic archebacteria that use anaerobic respiration for
ATP synthesis. Methanogens use CO2 taken up from their growth environment as the carbon
substrate for growth. They use some CO2 as the ultimate oxidizing agent of an electron transport
chain which, by a chemiosmotic mechanism, maintains a transmembrane electrochemical ion
gradient which powers ATP production. Methanogens use this hydrogen and this process
maintains a lowered hydrogen partial pressure in the reticulo-rumen. Some of the hydrogen
producing heterotrophic microorganisms show altered patterns of metabolism because of
methanogen usage of the hydrogen they produce. Methanogens affect the growth of some but not
all hydrogen producing species of microorganism in the reticulo-rumen. The equation shows the
reduction of CO2 by H2 to produce methane. This redox reaction sustains anaerobic respiration
which allows the production of ATP. The methane produced by reduction of the carbon dioxide
is lost from the reticulorumen by eructation. It is a waste of feed carbon because the rumen does
not have methanotrophic bacteria and the host ruminant cannot utilize this gas.

FACTORS AFFECTING ANAEROBIC PROCESS

The successful operation of anaerobic reactor depends on maintaining the environmental factors
close to the comfort of the microorganisms involved in the process. They are as follows
1. Temperature: Anaerobic processes like other biological processes operate in certain
temperature ranges. Mesophilic (25-450C) and thermophilic (45-650C) anaerobic digestion are
commonly applied in the field. Most full-scale anaerobic digesters are operated at mesophilic
temperature. Since wastewater and bio solids is discharged at relatively low temperature (e.g., 18
0C), recent research toward anaerobic treatment under psychrophilical condition becomes
attractive. For instance, microbial communities involved in digestion are sensitive to temperature
changes. The rate of anaerobic degradation of organic substrates generally increases in the order
of psychrophilic, mesophilic and thermophilic digestion.
2. pH Control: pH is an important factor for keeping functional anaerobic digestion. A typical
pH is in the range of 6.5-7.6. The accumulation of intermediate acids leads to pH drop during
fermentation. In order to maintain stable operation, it is necessary to add bicarbonate or
carbonate as an alkalinity buffer to neutralize volatile fatty acids and carbon dioxide.
3. Nutrients: Macronutrients are the elements that the cellular material of the anaerobic
microorganisms comprises, including hydrogen, nitrogen, oxygen, carbon, sulfur, phosphorus,
potassium, calcium, magnesium and iron. Normally, anaerobic microorganisms require these
elements presented with a concentration around 10 -4 M. In addition to the micronutrients, a
number of other elements, such as Ni and Co must be present in small amount, i.e. below10 -4
M. This is because that these elements are important for the growth of anaerobic organisms. For
example, Ni is necessary for activating factor F 430 , which is a co factor involved in
methanogenesis. But it can be inhibitory for fermentative as well as methanogens if it is present
in high concentration.
4. Toxicity: Besides ammonia and nitrate/nitrite, heavy metals, such as Zn, Cu and Cd can be
toxic to acidogenic bacteria. However, many of these elements and compounds can be tolerated
in relatively high concentration due to absorption in inert material contained in the reactor.
5. Retention Time: For the CSTR reactors, which are the most prevailingly used types of
reactors, hydraulic and solid retention time is the same. Retention time is an important
operational parameter that is easy to operate and control. Tremendous efforts have been put into
the research of the effect of retention time on anaerobic digestion. Biologically, only those who
are doubling time are shorter than the retention time can be kept in the reactor, so retention time
is one of the best parameter to be manipulated for separating and enriching different groups of
the microbes involved in the anaerobic process. Also, retention time determines the time that
substrates can be attacked by the enzymes in the reactor.
 6. Feeding Strategy: Practically, anaerobic reactors treating sewage sludge in wastewater
treatment plants are fed semi-continuously instead of continuously. Feeding frequency
determines the ratio of food to microbe (F/M) when the retention time and the working volume
have been fixed. Normally, the ratio can be satisfied so that there is no negative effect on the
stability and on the performance of the anaerobic reactors.
7. Agitation Strategy: It is normally believed that agitation is necessary to help the diffusion of
substrate and increase their contacts with the microbes, especially when raw sludge is
intermittently fed into the reactor. Agitation strategy can affect anaerobic digestion of sewage
sludge and optimum agitation strategy should be found. In addition, it was also found that
mixing levels might be used as an operational tool to stabilize unstable anaerobic reactor.

TYPES OF ANAEROBIC REACTORS

There are five principal process variants which are proper in anaerobic wastewater treatment.
These are as follows:

Anaerobic Filter Reactor: The anaerobic filter is similar to a trickling filter in that a biofilm is
generated on media. The bed is fully submerged and can be operated
either upflow or down flow. As wastewater flows through the filter, particles are trapped
and organic matter is degraded by the active biomass that is attached to the surface of
the filter material. With this technology, suspended solids and BOD removal can be as high as
90%, but is typically between 50% and 80%. Nitrogen removal is limited and normally does not
exceed 15% in terms of total nitrogen (TN). Anaerobic filters are usually operated in upflow
mode because there is less risk that the fixed biomass will be washed out. The water level should
cover the filter media by atleast 0.3 m to guarantee an even flow regime. The hydraulic retention
time (HRT) is the most important design parameter influencing filter performance. An HRT of
12 to 36 hours is recommended. The ideal filter should have a large surface area for bacteria to
grow, with pores large enough to prevent clogging. The surface area ensures increased contact
between the organic matter and the attached biomass that effectively degrades it. Ideally, the
material should provide between 90 to 300 m2 of surface area per m3 of occupied reactor
volume. Typical filter material sizes range from 12 to 55 mm in diameter. Materials commonly
used include gravel, crushed rocks or bricks, cinder, pumice, or specially formed plastic pieces,
depending on local availability.

Anaerobic Contact Process Reactor: This process can be considered as an anaerobic activated
sludge because sludge is recycled from a clarifier or separator to the reactor. Since the material
leaving the reactor is a gas-liquid-solid mixture, a vacuum Degasifier is required to separate the
gas and avoid floating sludge in the clarifier. Here a set of reactors are created in series, often
with recycling. This recycled material is pumped up into the bottom of the first reactor, an
upflow reactor. The upflow anaerobic process is a large reactor which allows the waste to flow
up from the bottom and separates the waste into 3 zones. At the very top is the biogas zone
where the gas is collected. Bacteria digest waste in the lowest portion of the upflow reactor; the
bioreactor zone. In between these two stages is the clarifier zone where the which exports the
stabilized waste.

Fluidized Bed Reactor: This reactor consists of a sand bed on which the biomass is grown.
Since the sand particles are small, a very large biomass can be developed in a small volume of
reactor. In order to fluidize the bed, a high recycle is required. In this type of reactor, a fluid (gas
or liquid) is passed through a granular solid material (usually a catalyst possibly shaped as tiny
spheres) at high enough velocities to suspend the solid and cause it to behave as though it were a
fluid. This process, known as fluidization, imparts many important advantages to the FBR. As a
result, the fluidized bed reactor is now used in many industrial applications.

Upflow Anaerobic Sludge Blanket Reactor: Under proper conditions anaerobic sludge will
develop as high density granules. These will form a sludge blanket in the reactor. The
wastewater is passed upward through the blanket. Because of its density, a high concentration of
biomass can be developed in the blanket. The UASB reactor is a methanogenic (methane-
producing) digester that evolved from the clarifier. A similar but variant technology to UASB is
the expanded granular sludge bed (EGSB) digester UASB uses an anaerobic process whilst
forming a blanket of granular sludge which suspends in the tank. Wastewater flows upwards
through the blanket and is processed (degraded) by the anaerobic microorganisms. The upward
flow combined with the settling action of gravity suspends the blanket with the aid of
flocculants. The blanket begins to reach maturity at around 3 months. Small sludge granules
begin to form whose surface area is covered in aggregations of bacteria. In the absence of any
support matrix, the flow conditions create a selective environment in which only those
microorganisms, capable of attaching to each other, survive and proliferate. Eventually the
aggregates form into dense compact biofilms referred to as "granules"

DESIGN OF ANAEROBIC REACTORS

Conditions for efficient anaerobic treatment –
 Avoid excessive air/O 2 exposure
 No toxic/inhibitory compounds present in the influent
 Maintain pH between 6.8 –7.2
 Sufficient alkalinity present (mainly bicarbonates)
 Low volatile fatty acids (VFAs)
 Temperature around mesophilic range (30-38 o C)
 Enough nutrients (N & P) and trace metals especially, Fe, Co, Ni, etc. COD - N: P =
350:7:1 (for highly loaded system) 1000:7:1 (lightly loaded system)
 SRT/HRT >>1 (use high rate anaerobic reactors)

DESIGN PARAMETERS
Anaerobic Filter: 
Width to diameter ratio of reactor = 2-6(usually)
Height of reactor = 2-12 m(usually)
Hydraulic retention time = 20-30 d(for domestic wastewater)
Volumetric loading = 0.2-0.8 kg COD/m3 –d
Specific area for the media = 100 m2 /m3 of volume
Fluidized Bed Reactor: 
Up –flow velocity =2.0 m/h 
Specific area of media =10000 m2 /m3 of media volume 
Void space =50% 
Organic loading =4-5 kg COD/m3 -d
Anaerobic Contact Process: 
Up-flow velocity =15-20 m/h  Rector depth =3-6 m 
Volumetric loading =10-30 kg COD/m3 .d 
Reactor biomass =15000-20000 mg MLVSS/L  HRT =3-6 h

UASB: 
Up-flow velocity =0.5-0.9 m/h 
Volumetric loading =6-20 kg COD/m3 .d 
HRT =6-48 hr 
MLSS concentration (i) at the bottom of reactor =100000-150000 mg/l (ii) at the top of reactor
=5000-4000 mg/l 
Reactor depth = 3-5 m (for domestic water) 
Biomass production =0.2-0.5 m3 /kg of COD removed
ADVANTAGES OF ANAEROBIC PROCESS
• Less energy requirement as no aeration is needed.
• Energy generation in the form of methane gas.
• Less biomass (sludge) generation.
• Less nutrients (N & P) required.
• Application of higher organic loading rate.
• Space saving.
• Ability to transform several hazardous solvents.
LIMITATIONS OF ANAEROBIC PROCESS
• Long start-up time.
• Long recovery time.
• Specific nutrients/trace metal requirements.
• More susceptible to changes in environmental conditions.
• Effluent quality of treated wastewater.
• Treatment of high protein & nitrogen containing wastewater.