Types Of Fermentations

Fermentation types are classified into different classes based on various aspects like based on
feeding substrate to fermenter, based on need of supply of aeration, based on need of light etc.
1. Based on feeding substrate to fermenter
a. Batch fermentation
b. Continuous fermentation
c. Fed-batch fermentation.
a. In batch fermentation, every material for process of fermentation including substrate, inoculum
and all the process parameters are set and filled in a fermenter and the process is set on and until
the total process comes to an end neither substrate is added into fermenter nor product is taken out
of fermenter. Its a closed system.
b. In continuous fermentation, the substrate is added continuously to the fermenter at a fixed rate
which maintains the microbes at logarithmic growth phase and the products that are formed are
taken out simultaneously and here we find growth associated products.
c. In fed-batch mode we find both modes of operations of batch and continuous modes, where
substrate is added at fixed time intervals during the fermentation process.
2. Based on need of supply of aeration
a. aerobic fermentation,
b. anaerobic fermentation
a. Aerobic fermentations: even though called fermentation, many large-scale fermentation
processes are carried out in presence of aerobic conditions where, the contents present in fermenter
are agitated with the help of agitator and with the help of spargers by forcing sterilized air into the
fermenter.
b. Anaerobic fermentation: apart from intense need and presence of agitator and sparger to supply
aeration, rest of the configuration of the fermenter is as same as aerobic fermentation. But the
presence of agitator is made compulsion for the even distribution of temperature, pH, viscosity,
nutrients etc. along the medium in the fermenter. Anyways, micro aerophilic conditions to
anaerobic conditions are needed to initiate the growth of the microbe in the fermenter. 3. Based on
need of light
a. photo fermentation (only photosynthetic bacteria can undergo),
b. Dark fermentation.
a. Photofermentation: its a process of conversion of organic substances to other utilisable energy
compounds following a series of biochemical reactions carried out by a specific group of bacteria
named Photosynthetic bacteria, which only proceeds in the presence of light.
b. Dark fermentation: in every way dark fermentation is similar to that of photofermentation, but
in aspect of need of light, dark fermentation does not need any light to initiate the reactions and a
diversified group of microbes are involved in dark fermentation.

Design of a Fermentor
Fermentation
Fermentation can be defined as a metabolic process in which cheap raw materials such as sugar or
carbohydrates are converted into economically important products like acids, gases and alcohols
by micro-organism. This process is carried out in a equipment called as fermentor.
Fermentation Technology could be defined simply as the study of the fermentation process,
techniques and its application. Fermentation should not be seen merely as a process that is entirely
focused on the happenings occurring in the fermenter alone! There are many activities that occur
upstream leading to the reactions that occur within the bioreactor or fermenter, despite the
fermenter is regarded as the heart of the fermentation process.

Fermentation technology is the whole field of study which involves studying, controlling and
optimization of the fermentation process right up from upstream activities, mid stream and
downstream or post fermentation activities.

The study of fermentation technology requires essential inputs from various disciplines such as
biochemistry, microbiology, genetics, chemical and bioprocess engineering and even a scatter of
mathematics and physics.

Fermentation in terms of biochemistry and physiology:-

Fermentation is now defined as a process of energy generation by various organisms especially
microorganisms. The fermentation process showed unique characteristics by which it generates
energy in the absence of oxygen. The process of energy generation utilizes the use of substrate
level phosphorylation (SLP) which do not involved the use of electron transport chain and free
oxygen as the terminal electron acceptor.
Engineers definition of fermentation:-
It is only up to recently with the rise of industrial microbiology and biotechnology that
the definition of fermentation took a less specific meaning. Fermentation is defined more from
the point of view of engineers. They see fermentation as the cultivation of high amount of
microorganisms and biotransformation being carried out in special vessels called fermenter or
bioreactors.
Their definitions make no attempt to differentiate whether the process is aerobic or anaerobic.
Neither are they bothered whether it involves microorganisms or single animal or plant cells.
They view bioreactors as a vessel which is designed and built to support high concentration of
cells.
A bioreactor is a specially designed vessel which is built to support the growth of high
concentration of microorganisms. It must be so designed that it is able to provide the optimum
environments or conditions that will allow supporting the growth of the microorganisms.

Bioreactors are commonly cylindrical vessels with hemispherical top and/or bottom, ranging
in size from some liter to cube meters, and are often made of stainless steel and glass.

The difference between a bioreactor and a typical composting system is that more parameters
of the composting process can be measured and controlled in bioreactors.

The sizes of the bioreactor can vary over several orders of magnitudes. The microbial cell

(few mm3), shake flask (100-1000 ml), laboratory fermenter (1 – 50 L), pilot scale (0.3 –

10m3) to plant scale (2 – 500 m3) are all examples of bioreactors.

The design and mode of operation of a fermenter mainly depends on the production organism,
the optimal operating condition required for target product formation, product value and
scale of production. The design also takes into consideration the capital investment and running
cost.
 • Large volume and low value products like alcoholic beverages need simple fermenter
and do not need aseptic condition.
• High value and low volume products require more elaborate system of operation
and aseptic condition.

Bioreactors differ from conventional chemical reactors in that they support and control
biological entities. As such, bioreactor systems must be designed to provide a higher degree of
control over process upsets and contaminations, since the organisms are more sensitive and less
stable than Chemicals. Biological organisms, by their nature, will mutate, which may alter the
biochemistry of the bioreaction or the physical properties of the organism. Analogous to
heterogeneous catalysis, deactivation or mortality occur and promoters or coenzymes
influence the kinetics of the bioreaction. Although the majority of fundamental bioreactor
engineering and design issues are similar, maintaining the desired biological activity and
eliminating or minimizing undesired activities often presents a greater challenge than
traditional chemical reactors typically require.

Other key differences between chemical reactors and bioreactors are selectivity and rate. In
bioreactors, higher selectivity — that is, the measure of the system’s capability for producing
the preferred product (over other outcomes) — is of primary importance. In fact,
selectivity is especially important in the production of relatively complex molecules such
as antibiotics, steroids, vitamins, proteins and certain sugars and organic acids. Frequently,
the activity and desired selectivity occur in a substantially smaller range of conditions
than are present in conventional chemical reactors. Further, deactivation of the biomass
often poses more severe consequences than a chemical upset.
The Designing of a Bioreactor also has to take into Considerations the Unique
Aspects of Biological Processes

a. The concentrations of starting materials (substrates) and products in the reaction mixture are
frequently low; both the substrates and the products may inhibit the process. Cell growth, the
structure of intracellular enzymes, and product formation depend on the nutritional needs of
the cell (salts, oxygen) and on the maintenance of optimum biological conditions (temperature,
concentration of reactants, and pH) within narrow limits.
b. Certain substances inhibitors effectors, precursors, metabolic products influence the rate and
the mechanism of the reactions and intracellular regulation.
c. Microorganisms can metabolize unconventional or even contaminated raw materials
(cellulose, molasses, mineral oil, starch, wastewater, exhaust air, biogenic waste), a process
which is frequently carried out in highly viscous, non-Newtonian media.
d. In contrast to isolated enzymes or chemical catalysts, microorganisms adapt the structure and
activity of their enzymes to the process conditions, whereby selectivity and productivity
can change. Mutations of the microorganisms can occur under sub optimal biological
conditions.
e. Microorganisms are frequently sensitive to strong shear stress and to thermal and
chemical influences.
f. Reactions generally occur in gas-liquid -solid systems, the liquid phase usually being
aqueous.
g. The microbial mass can increase as biochemical conversion progresses. Effects such as
growth on the walls, flocculation, or autolysis of microorganisms can occur during the reaction.
h. Continuous bioreactors often exhibit complicated dynamic
behaviour.

Requirements of Bioreactors
Due to above mentioned demands made by biological systems on their environment, there is no
universal bioreactor. However, the general requirements of the bioreactor are as follows:

1. The vessel should be robust and strong enough to withstand the various treatments required
such as exposure to high heat, pressure and strong chemicals and washings and cleanings.
2. The vessel should be able to be sterilized and to maintain stringent aseptic conditions over
long periods of the actual fermentation process.
3. The vessel should be equipped with stirrers or mixers to ensure mass transfer processes occur
efficiently.
4. It should have sensors to monitor and control the fermentation process.
5. It should be provided with inoculation point for aseptic transfer in inoculum.
6. Sampling valve for withdrawing a sample for different tests.
7. Baffles should be provided in case of stirred fermenter to prevent vertex formation.
8. It should be provided with facility for intermittent addition of an antifoam agent.
9. In case of aerobic submerged fermentation, the tank should be equipped with the
aerating device.
10. Provision for controlling temperature and pH of fermentation medium.
11. Man hole should be provided at the top for access inside the fermenter for different
purposes.

Fermenter Design
What should be the basic points of consideration while designing a fermenter?
ƒ Productivity and yield
ƒ Fermenter operability and reliability
ƒ Product purification
ƒ Water management
ƒ Energy requirements
ƒ Waste treatment
 Few Significant things of concern that should be taken into account while
designing a fermenter:
ƒ Design in features so that process control will be possible over reasonable
ranges of process variables.
ƒ Operation should be reliable
ƒ Operation should be contamination free.
ƒ Traditional design is open cylindrical or rectangular vessels made from wood or
stone.
ƒ Most fermentation is now performed in close system to avoid contamination.
ƒ Since the fermenter has to withstand repeated sterilization and cleaning, it
should be constructed from non-toxic, corrosion-resistant materials.
ƒ Small fermentation vessels of a few liters capacity are constructed from glass
and/or stainless steel.
ƒ Pilot scale and many production vessels are normally made of stainless steel with
polished internal surfaces.
ƒ Very large fermenter is often constructed from mild steel lined with glass or
plastic, in order to reduce the cost.
ƒ If aseptic operation is required, all associated pipelines transporting air, inoculum
and nutrients for the fermentation need to be sterilizable, usually by steam.
ƒ Most vessel cleaning operations are now automated using spray jets, which are
located within the vessels. They efficiently disperse cleaning fluids and this cleaning
mechanism is referred to as cleaning-in-place CIP.
Associated pipe work must also be designed to reduce the risk of microbial contamination.
There should be no horizontal pipes or unnecessary joints and dead stagnant spaces
where material can accumulate; otherwise this may lead to ineffective sterilization.
Overlapping joints are unacceptable and flanged connections should be avoided as vibration
and thermal expansion can result in loosening of the joints to allow ingress of microbial
contaminants.
ƒ Butt welded joints with polished inner surfaces are preferred.
 Normally, fermenters up to 1000 L capacity have an external jacket, and larger vessels have
internal coils. Both provide a mechanism for vessel sterilization and temperature control
during the fermentation.
Other features that must be incorporated are pressure gauges and safety pressure valves, which
are required during sterilization and operation. The safety valves prevent excess pressurization,
thus reducing potential safety risks. They are usually in the form of a metal foil disc held in a
holder set into the wall of the fermenter. These discs burst at a specified pressure and present
a much lower contamination risk than spring-loaded valves.
For transfer of media pumps are used. However pumps should be avoided if aseptic
operation is required, as they can be a major source of contamination. Centrifugal pumps may
be used, but their seals are potential routes for contamination. These pumps generate high
shear forces and are not suitable for pumping suspensions of shear sensitive cells. Other
pumps used include magnetically coupled, jet and peristaltic pumps.
Alternate methods of liquid transfer are gravity feeding or vessel pressurization. In fermentations
operating at high temperatures or containing volatile compounds, a sterilizable condenser may
be required to prevent evaporation loss. For safety reasons, it is particularly important to contain
any aerosols generated within the fermenter by filter- sterilizing the exhaust gases.
Also, fermenters are often operated under positive pressure to prevent entry of
contaminants.

2. Considerations that improve productivity of bioreactor: -

2.1 Material of construction: -
2.1.1 Laboratory scale bioreactor:
In fermentation with strict aseptic requirements it is important to select materials that
can withstand repeated sterilization cycles. On a small scale, it is possible to use glass and/or
stainless steel.
Glass is useful because it gives smooth surfaces, is non-toxic, corrosion proof and it is usually
easy to examine the interior of vessel. The glass should be 100% borosilicate, e.g. Pyrex® and
Kimax®.
The following variants of the laboratory bioreactor can be made:
1. Glass bioreactor (without the jacket) with an upper stainless steel lid.
2. Glass bioreactor (with the jacket) with an upper stainless steel lid.
3. Glass bioreactor (without the jacket) with the upper and lower stainless steel lids.
4. Two-part bioreactor - glass/stainless steel.
The stainless steel part has a jacket and ports for electrodes installation.
5. Stainless steel bioreactor with peepholes.
Vessels with two stainless steel plates cost approximately 50% more than those with just a
top plate. The geometrical term of the bioreactors are shown in the figure on the next page.

2.3. Agitation: -

2.3.1 The agitator (impeller).

The agitator is required to achieve a number of mixing objective.
¾ Bulk fluid and gas-phase mixing,
¾ Air dispersion,
¾ Oxygen transfer,
¾ Heat transfer,
¾ Suspension of solid particles and maintain a uniform environment throughout the
vessel contents.
¾ Enhancement of mass transfer between dispersed phase
Bulk mixing and micro mixing both are influenced strongly by impeller type, broth rheology,
and tank geometry and internals.
Impellers used bioreactors are:

Rushton disc turbines, vaned discs, open turbines of variable pitch and propeller. The disc
turbine consists of a disc with a series of rectangular vanes set in vertical plane around the
circumstances and vaned disc has a series of rectangular vanes attached vertically to the
underside. Air from the sparger hits the underside of the disc and is displaced towards the vanes
where the air bubbles are broken up into smaller bubbles. The vanes of variable pitch open
turbine and the blade of marine impellers are attached directly to a boss on the agitator
shaft. In this case air bubbles do not initially hit any surface before dispersion by the vanes
or blades.
 The Rushton disc turbine is the one used most often for highly aerobic fermentations, because
it has among the highest power draws of any the commercially available impellers, and it is
better characterized than others; hence, its behaviour is easier to predict. Rushton disc
turbine of one third of the fermenter diameter has been considered the optimum design for use
in fermentation processes. Disc turbine is most suitable in a fermenter since it can break
up a fast air stream without itself becoming flooded in air bubbles. A marine propeller is an
axial flow impeller which provides good top-to-bottom mixing. It is low power device does not
provide large oxygen-transfer rates.

The propeller and the open turbine flood when superficial velocity (Vs) exceeds 21 m h-1 ,

whereas the flat blade turbine can tolerate Vs of 120 m h-1 before being flooded, when two
sets are used on the same shaft. Besides this, propeller is also less efficient in breaking up the
bubbles and the flow it produces is axial rather than the radial. One of major drawback of
Rushton disc turbine is that it
provides very axial flow, resulting in poor overall top-to-bottom mixing. In addition, agitation
intensity decrease with distance from the impeller, and this decrease can become more
pronounced for viscous, pseudoplastic broths
2.3.2 Various impellers use in bioreactors with their flow patterns.
Flat blade disk 45° Flat blade disk Curved blade disk
turbine turbine turbine
 Pitched blade turbine Marine propeller Intermig

3 segment blade
curved blade turbine Large pitch blade
impeller
impeller

Temperature Control
During the fermentation process heat can be produced mainly in two ways, firstly microbial
biochemical reactions and secondly mechanical agitation. In case of fermentation, a temperature
control helps to control the temperature at the optimum level by removing or providing heat. In
small scale production vessel the amount of produced heat is negligible. Therefore, extra heat is
provided by hot bath or internal heat coil or heating jacket with a water circulation system or
silicon heating jacket. The silicon heating jacket consists of silicon rubber mats with heating wires
and it is wrapped around the fermenter. In the case of pilot-scale fermenters, it is not possible to
use silicon jackets due to large size. In such cases, an internal heating coil is used for providing
extra heat while cold water circulation helps to remove excess heat.
Stirrer glands and bearings
The most important factor of designing a fermenter is to maintain aseptic conditions inside the
vessel. It is highly challenging in the case of pilot-scale fermenters. Therefore stirrer shafts are
required. These stirrer shafts play an important role to seal the openings of a bioreactor. As a result,
it restricts the entry of air from outside. There are several types of seals used for this purpose,
which are following

The Stuffing Box: The shafted is sealed by several layers of packing rings of asbestos or cotton
yarn which is pressed against the shaft by gland follower. At high stirrer speeds, the packing wears
quickly and excessive pressure may need to ensure the tightness of fit. The packing may be difficult
to sterilize properly because of unsatisfactory heat penetration and it is necessary to check and
replace the packing rings regularly.
The Mechanical Seal: It is used in both small scale and large scale fermenters. The seal is divided
into two parts, first is the stationary bearing housing and the second rotates on the shaft. These two
parts are pressed together by springs. Steam condensate is used to lubricate and cool the seals
during operation and provides protection against the contamination.
Magnetic Drives: This type of seals helps to counter the problem originated by the impeller shaft
which is going through the top or bottom of the fermenter plate. The magnetic drive is made up of
two magnets one is driving and one driven. The driven magnet held in bearings in housing on the
outside of the head plate and connected to a drive shaft. The internal driven magnet is placed on
one end of the impeller shaft and held in bearings in a suitable housing on the inner surface of the
head plate. When multiple ceramic magnets have been used it has been possible to transmit power
across a gap of 16mm. Using this drive water can be stirred in baffled vessels up to 300 dm3
capacity at speeds of 300 to 2000 rpm.

Baffles
There are four baffles that are present inside of an agitated vessel to prevent a vortex and improve
aeration efficiency. Baffles are made up of metal strips roughly one-tenth of the vessel diameter
and attached to the wall. The agitation effect is slightly increased with wider baffles but drops
sharply with narrower baffles. After installation of the baffle there a gap between them and the
vessel wall which facilitates scouring action around the baffles and minimizes microbial growth
on the baffles and the fermenter wall. Baffles are often attached to cooling coils to increase the
cooling capacity of the fermenter.

The aeration system (sparger)

A sparger is a device that introduces air into the liquid medium in a fermenter. There are three
main types of fermenter used in industrial-scale bioreactors such as
Porous Sparger: It is made up of sintered glass, ceramics or metals’ and are mostly used in
laboratory-scale bioreactors. As it introduces air inside a liquid medium, bubbles are formed.
These bubbles are always 10 to 100 times larger than the pore size of the aerator. The air pressure
is generally low in these devices and a major disadvantage of using porous sparger is that microbial
growth may occur on the pores which hamper the airflow.
Orifice Sparger: These are used in small stirred fermenters where perforated pipes are used and
attached below the impeller in the form of a ring. The air holes are mostly drilled under the surface
of the tubes. Orifice spargers were used to a limited extent in yeast manufacture, effluent treatment
and production of single-cell proteins.
Nozzle Sparger: This is used in industrial-scale fermenters. The main characteristic of this kind of
sparger is that it contains a single open or partially closed pipe as an air outlet. The pipe needs to
be positioned below the impeller. The design helps to overcome troubles related to sparger
blockage.
pH control sensors
All types of fermenters are attached with a pH control sensor which consists of a pH sensor and a
port to maintain the pH inside of the fermenter. pH alteration can lead to death of the organism
which leads to product loss. Therefore, it is a crucial instrument for a fermenter and needs to be
checked regularly.

Media
A growth medium or culture medium is a solid, liquid or semi-solid designed to support the growth
of a population of microorganisms or cells via the process of cell proliferation, or small plants like
the moss Physcomitrella patens. Different types of media are used for growing different types of
cells.
The two major types of growth media are those used for cell culture, which use specific cell types
derived from plants or animals, and microbiological culture, which are used for growing
microorganisms, such as bacteria or fungi. The most common growth media for microorganisms
are nutrient broths and agar plates; specialized media are sometimes required for microorganism
and cell culture growth. Some organisms, termed fastidious organisms, require specialized
environments due to complex nutritional requirements. Viruses, for example, are obligate
intracellular parasites and require a growth medium containing living cells.
Types

US Food and Drug Administration scientist tests for Salmonella

The most common growth media for microorganisms are nutrient broths (liquid nutrient medium)
or lysogeny broth medium. Liquid media are often mixed with agar and poured via a sterile media
dispenser into Petri dishes to solidify. These agar plates provide a solid medium on which microbes
may be cultured. They remain solid, as very few bacteria are able to decompose agar (the exception
being some species in the genera: Cytophaga, Flavobacterium, Bacillus, Pseudomonas, and
Alcaligenes). Bacteria grown in liquid cultures often form colloidal suspensions.

The difference between growth media used for cell culture and those used for microbiological
culture is that cells derived from whole organisms and grown in culture often cannot grow without
the addition of, for instance, hormones or growth factors which usually occur in vivo. In the case
of animal cells, this difficulty is often addressed by the addition of blood serum or a synthetic
serum replacement to the medium. In the case of microorganisms, no such limitations exist, as
they are often unicellular organisms. One other major difference is that animal cells in culture are
often grown on a flat surface to which they attach, and the medium is provided in a liquid form,
which covers the cells. In contrast, bacteria such as Escherichia coli may be grown on solid or in
liquid media.

An important distinction between growth media types is that of defined versus undefined media.
A defined medium will have known quantities of all ingredients. For microorganisms, they consist
of providing trace elements and vitamins required by the microbe and especially defined carbon
and nitrogen sources. Glucose or glycerol are often used as carbon sources, and ammonium salts
or nitrates as inorganic nitrogen sources. An undefined medium has some complex ingredients,
such as yeast extract or casein hydrolysate, which consist of a mixture of many chemical species
in unknown proportions. Undefined media are sometimes chosen based on price and sometimes
by necessity – some microorganisms have never been cultured on defined media.

A good example of a growth medium is the wort used to make beer. The wort contains all the
nutrients required for yeast growth, and under anaerobic conditions, alcohol is produced. When
the fermentation process is complete, the combination of medium and dormant microbes, now
beer, is ready for consumption. The main types are

Cultural media
Minimal media
Selective media
Differential media
Transport media
Indicator media
Culture media
Culture media contain all the elements that most bacteria need for growth and are not selective, so
they are used for the general cultivation and maintenance of bacteria kept in laboratory culture
collections.
Physcomitrella patens plants growing axenically on agar plates (Petri dish, 9 cm diameter)
An undefined medium (also known as a basal or complex medium) contains:
a carbon source such as Glucose water various salts a source of amino acids and nitrogen (e.g.,
beef, yeast extract)
This is an undefined medium because the amino-acid source contains a variety of compounds with
the exact composition being unknown.
A defined medium (also known as chemically defined medium or synthetic medium) is a medium
in which all the chemicals used are known no yeast, animal, or plant tissue is present
Some examples of nutrient media include:
Plate count agar
Nutrient agar
Trypticase soy agar
Minimal media
A defined medium that has just enough ingredients to support growth is called a "minimal
medium". The number of ingredients that must be added to a minimal medium varies enormously
depending on which microorganism is being grown. Minimal media are those that contain the
minimum nutrients possible for colony growth, generally without the presence of amino acids, and
are often used by microbiologists and geneticists to grow "wild-type" microorganisms. Minimal
media can also be used to select for or against recombinants or exconjugants.

Minimal medium typically contains:

a carbon source, which may be a sugar such as glucose, or a less energy-rich source such as
succinate
various salts, which may vary among bacteria species and growing conditions; these generally
provide essential elements such as magnesium, nitrogen, phosphorus, and sulfur to allow the
bacteria to synthesize protein and nucleic acids
water
Supplementary minimal media are minimal media that also contains a single selected agent,
usually an amino acid or a sugar. This supplementation allows for the culturing of specific lines of
auxotrophic recombinants.

Selective media

Blood-free, charcoal-based selective medium agar (CSM) for isolation of Campylobacter

Blood agar plates are often used to diagnose infection. On the right is a positive Staphylococcus
culture; on the left is a positive Streptococcus culture.
Selective media are used for the growth of only selected microorganisms. For example, if a
microorganism is resistant to a certain antibiotic, such as ampicillin or tetracycline, then that
antibiotic can be added to the medium to prevent other cells, which do not possess the resistance,
from growing. Media lacking an amino acid such as proline in conjunction with E. coli unable to
synthesize it were commonly used by geneticists before the emergence of genomics to map
bacterial chromosomes.

Selective growth media are also used in cell culture to ensure the survival or proliferation of cells
with certain properties, such as antibiotic resistance or the ability to synthesize a certain metabolite.
Normally, the presence of a specific gene or an allele of a gene confers upon the cell the ability to
grow in the selective medium. In such cases, the gene is termed a marker.

Selective growth media for eukaryotic cells commonly contain neomycin to select cells that have
been successfully transfected with a plasmid carrying the neomycin resistance gene as a marker.
Gancyclovir is an exception to the rule, as it is used to specifically kill cells that carry its respective
marker, the Herpes simplex virus thymidine kinase.
Four types of agar plate demonstrating differential growth depending on bacterial metabolism
Examples of selective media include:
Eosin methylene blue contains dyes that are toxic for Gram-positive bacteria. It is the selective
and differential medium for coliforms.
YM (yeast extract, malt extract agar) has a low pH, deterring bacterial growth.
MacConkey agar is for Gram-negative bacteria.
Hektoen enteric agar is selective for Gram-negative bacteria.
HIS-selective medium is a type cell culture medium that lacks the amino acid histidine.
Mannitol salt agar is selective for Gram-positive bacteria and differential for mannitol.
Xylose lysine deoxycholate is selective for Gram-negative bacteria.
Buffered charcoal yeast extract agar is selective for certain Gram-negative bacteria, especially
Legionella pneumophila.
Baird–Parker agar is for Gram-positive staphylococci.
Sabouraud's agar is selective to certain fungi due to its low pH(5.6) and high glucose
concentration(3-4%)
DRBC (Dichloran Rose Bengal Chloramphenicol agar) is a selective medium for the enumeration
of moulds and yeasts in foods. Dichloran and rose bengal restricts the growth of mould colonies
thus preventing overgrowth of luxuriant species and assisting accurate counting of colonies.
Differential media
UTI Agar is a chromogenic medium for differentiation of main microorganisms that cause urinary
tract infections (UTIs).
Differential or indicator media distinguish one microorganism type from another growing on the
same medium. This type of media uses the biochemical characteristics of a microorganism
growing in the presence of specific nutrients or indicators (such as neutral red, phenol red, eosin
y, or methylene blue) added to the medium to visibly indicate the defining characteristics of a
microorganism. These media are used for the detection of microorganisms and by molecular
biologists to detect recombinant strains of bacteria.

Examples of differential media include:

Blood agar (used in strep tests) contains bovine heart blood that becomes transparent in the
presence of β-hemolytic organisms such as Streptococcus pyogenes and Staphylococcus aureus.
Eosin methylene blue is differential for lactose fermentation.
Granada medium is selective and differential for Streptococcus agalactiae (group B streptococcus)
which grows as distinctive red colonies in this medium.
MacConkey agar is differential for lactose fermentation.
Mannitol salt agar is differential for mannitol fermentation.
X-gal plates are differential for lac operon mutants.
Transport media
Transport media should fulfill these criteria:
Temporary storage of specimens being transported to the laboratory for cultivation
Maintain the viability of all organisms in the specimen without altering their concentration
Contain only buffers and salt
Lack of carbon, nitrogen, and organic growth factors so as to prevent microbial multiplication
Transport media used in the isolation of anaerobes must be free of molecular oxygen.
Examples of transport media include:
Thioglycolate broth is for strict anaerobes.
Stuart transport medium is a non-nutrient soft agar gel containing a reducing agent to prevent
oxidation, and charcoal to neutralize.
Certain bacterial inhibitors are used for gonococci, and buffered glycerol saline for enteric bacilli.
Venkataraman Ramakrishna (VR) medium is used for V. cholerae

Enriched media
Enriched media contain the nutrients required to support the growth of a wide variety of organisms,
including some of the more fastidious ones. They are commonly used to harvest as many different
types of microbes as are present in the specimen. Blood agar is an enriched medium in which
nutritionally rich whole blood supplements the basic nutrients. Chocolate agar is enriched with
heat-treated blood (40–45 °C), which turns brown and gives the medium the color for which it is
named
 SCHOOL OF BIO AND CHEMICAL ENGINEERING
DEPARTMENT OF BIOTECHNOLOGY

UNIT – III– Bioprocess Engineering -1 – SBT1301