Milk and

Dairy Products
 Milk Products -Yogurt
• Yogurt is a mixture of milk (whole, reduced-fat, low fat or
non fat) and cream fermented by a culture of lactic acid-
producing bacteria, Lactobacillus bulgaricus and
Streptococcus thermophilus.

• Other bacteria (e.g., acidophilus) and other strains of the

above bacteria may be added to the culture.
• Yogurt contains at least 3.25% milk fat and 8.25% solids-
not-fat. The mixture of dairy products and optional
ingredients, except bulky flavorings, must be pasteurized
or ultra pasteurized.
• The milk in most yogurts is also homogenized. Some
yogurts carry a seal on the label indicating that the yogurt
contains a significant level of
live, active cultures.
 Milk Products - CHEESE
Cheese is made by coagulating or curdling milk, stirring and
heating the curd, draining off the whey (the watery part of
milk),collecting and pressing the curd,and in some cases,
ripening.
Cheese can be made from whole, 2% lowfat, 1% lowfat or
fat-free milk, or combinations of these milks.
Starter Cultures
Starter cultures have crucial roles to play during
all phases of the cheese making and maturation
process.

Starter cultures, consisting of bacterial cultures, including

Lactococcus lactis, are added to start the cheese making
process. As the culture grows in the milk, it converts lactose
to lactic acid. This ensures the correct pH for coagulation and
influences the final moisture content, texture and flavor of
the product.
 Types of Cheese
 More than 400 different varieties of cheese are available.
 Cheeses are categorized in several ways:
 natural versus process cheeses,
 unripened versus ripened and
 soft versus hard.
 Many cheeses are named for their place of origin,
 such as Cheddar cheese, which originated in Cheddar, England.

Natural cheeses are often categorized according to their moisture or degree

of softness or hardness
• Unripened cheeses are made by coagulating milk proteins (casein) with
acid.Examples include soft cheeses like cream cheese, cottage cheese and
Neufchatel.
• Ripened cheeses are made by coagulating milk proteins with enzymes
(rennet) and culture acids.
 These cheeses are then ripened (aged) by bacteria or mold. Cheddar,
Swiss, Colby, brick and Parmesan are some examples of bacteria-ripened
cheeses.
Natural cheeses include:
 Soft Cheeses: Brie, Camembert, ricotta, cottage
 Semi-Soft Cheeses: Blue, brick, feta, Havarti, Monterey
Jack, mozzarella, Meunster, provolone
 Hard Cheeses: Cheddar, Colby, Edam, Gouda, Swiss
 Very Hard Cheese: Parmesan, Romano

 Process Cheeses. These cheeses are made by blending

one or more natural cheeses, heating and adding
emulsifying salts. Process cheeses contain more
moisture than natural cheeses.
 Pasteurized process cheeses include American cheese,
cheese spreads and cheese foods.
 Cold-pack cheese is a blend of natural cheeses
processed without heat.
 Bread is a staple food prepared from a dough of flour and water, usually
by baking.
 Throughout recorded history, it has been a prominent food in large parts of
the world.
 It is one of the oldest human-made foods, having been of significant
importance since the dawn of agriculture, and plays an essential role in
both religious rituals and secular culture.
 Bread may be leavened by naturally occurring microbes, chemicals,
industrially produced yeast, or high-pressure aeration which creates the gas
bubbles that fluffs up bread.
 In many countries, commercial bread often contains additives to
 improve flavor,
 texture,
 color,
 shelf life,
 nutrition, and ease of production.
 Chapter 6
Application of microbe in agricultural
biotechnology
 Biofertilizers

• Biofertilizers are the microbial inoculants which can be

usually defined as a preparation containing live or dormant
cells of efficient strains of
– nitrogen fixing,
– phosphate solubilizing,
– and cellulytic microorganisms, etc.
• In contrast to chemical fertilizers, biofertilizers are viable
microorganisms which are not the source of nutrients but
provide help to plants in accessing the nutrient availability in
rhizospheric region.
• Several microorganisms are commonly used as biofertilizers
including nitrogen-fixing soil bacteria (Azotobacter,
Rhizobium), nitrogen-fixing
 phosphate-solubilizing bacteria (Pseudomonas sp.),
phytohormone (auxin)-producing bacteria and
cellulolytic microorganisms are also used as biofertilizer
• These microbial formulations are used to enhance
certain microbial process to increase the availability of
nutrients in a form which can be assimilated by plant.
• Biofertilizers are
• low-cost,
• renewable sources of plant nutrients.
• These are the strains of beneficial soil microorganisms
which are cultured and packed in suitable carrier in
laboratory.
• A carrier is a material, such as peat, lignite powder,
vermiculite, clay, talc, rice bran, seed, charcoal, soil,
rock phosphate pellet, paddy straw compost,
wheat bran, or a mixture of such materials, etc. which
provides better shelf life to biofertilizer formulation.
Biofertilizers (microbial inoculants)
• Biofertilizers = fertilizers from biological origin
• The use of chemical (synthetic) fertilizers is the common
practice to increase crop yields.
• Beside the cost factor, the use of synthetic fertilizers is
associated with environmental pollution.
• To reduce the impact of excess chemical fertilizers in the field
of agriculture the biofertilizer is being considered as a potential
tool; biologically fixed nitrogen is such a source which can
supply an adequate amount of Nitrogen to plants and other
nutrients to some extent. Many free living and symbiotic
bacteria, which fix atmospheric Nitrogen are used as
biofertiliser material as a substitute for Nitrogen fertilizer. In
general two types of biofertiliser are used
 Bacterial Biofertilizer
 Algal Biofertilizer 11
 Generally, the term biofertilizers is used to refer to the
nutrient inputs of biological origin to support plants
growth.
 This can be achieved by the addition of microbial
inoculants as a source of biofertilizers.
Biofertilizers broadly includes the following categories
1. Symbiotic nitrogen fixers
e.g. Diazotrophic microorganisms including Rhizobium sp.
2. Asymbiotic nitrogen fixers
These microorganisms can directly convert the gaseous
nitrogen to nitrogen rich compounds when these
asymbiotic nitrogen fixers die,,
and thus serves as biofertilizers. e.g. Azobactor sp. The
most common one is blue green algae (cyanobacteria) 12
3. Phosphate solubilizing bacteria
Certain bacteria are capable of converting non-
valuable inorganic phosphorous present in the soil
to utilizable form of phosphate. E.g. Bacillus sp.
specially thiobacillus
4. Organic fertilizers
Includes several organic wastes, including animal
dung, urine, urban garbage, sewage, crop residue,
etc.
 What is bio-control?

• When we use natural enemies to reduce invasive species

populations, we refer to the natural enemies as
"biological control agents," or sometimes "biocontrol
agents."
• Biological control can be defined as the use of living
organisms to depress the population of a pest.
• However, biological control could be more accurately
called "biological suppression," i.e. reducing the
population of the target weed to an acceptable level.
• Often the goal of those who use biological control
agents on rangelands is to suppress the alien weeds and
at least partially restore the native plant community.
 Bio-pesticides Types:
Bio-Herbicides and Bio-Insecticides!
• Bio-pesticides are those biological agents that
are used for control of weeds, insects and
pathogens.
• The micro-organisms used as bio-pesticides
are viruses, bacteria, protozoa, fungi and
mites. Some of the bio-pesticides are being
used on a commercial scale.
 Bioinsecticides
• Bioinsecticides include bacteria, viruses, and fungi
that contrast to most synthetic insecticides as they can
be highly specific for certain pests
• may persist for a shorter time in the environment than
synthetic insecticides. ... Entomopathogenic viruses
are potentially important biological control agents
 How is Bioinsecticide produced?

• To develop a cost-effective process for the production

of Bacillus thuringiensis-based insecticide
• it is important to cultivate the bacterial strain in rich
medium to obtain the highest yields of spore-crystal
complexes. ...
• Only 78.3% of consumed glucose was converted into
spores and crystal protein
Bio-insecticides:
• Bio-insecticides are those biological agents that are
used to control harmful insects. They include the
following.
(a) Predators:
• Destructive insects or plant pests can be brought
under control through introduction of their natural
predators.
• The predators should be specific and unable to harm
the useful insects.
• Introduction of ladybugs (Lady Bird Beetles) and
Praying Mantis has been successful in combating
scale insects or aphids which feed on plant sap.
(b) Parasites and Pathogens:
• This is alternate biological control of plant pests
through the search of their natural parasites and
pathogens.
• They include viruses, bacteria, fungi and insect
parasitoids. Parasitoids are organisms that live as
parasites for some time (as early or larval stage) and
free living at other times, e.g., Trichogramma.
Nucleopolyhedrovirus (NPV) are species specific.
• For example, Bacillus thuringenesis (a bacterium) is
effective against the cabbage looper
 (C) Natural Insecticides
• They are insecticides and related pesticides which are
ob­tained from microbes and plants.
• A number of natural insecticides are available.
• The common ones include (i) Azadirachtin from
Margosa or Neem (Azadirachta indica). It occurs in
Margosa extract.
• Spray of the same keeps away the Japanese beetles
and other leaf eating pests because of the antifeedant
property of azadirachtin. (ii) Rotenones.
• They are powerful insecticides which are harmless to
warm blooded animals.
• Bioherbicides are biologically based control
agents useful for biological weed control.
• Hence, bioherbicides have been identified as a
significant biological control strategy.
• Bioherbicides have many advantages such
– as clearly defined for target weeds,
– no side effect on beneficial plants or human health,
– a lack of pesticide residue build-up in the
environment, and effectiveness for control of some
herbicide-resistant weed biotypes.
• Bioherbicides offer many advantages in comparison
with synthetic herbicides.
• They include:
• a high degree of specificity of target weed;
• no effect on non-target and beneficial plants or
man;
• absence of residue build-up in the environment;
and
• effectiveness for managing herbicide-resistant
(HR) weed populations
• Limitations in bioherbicide development can be
classified as either
• Environmental (temperature and, particularly,
humidity as major factors influencing the efficacy of
bioherbicides),
• Biological (mainly host variability and resistance),
• Or technological–commercial (mass production and
formulation,
• Land restoration can include the process of cleaning
up and rehabilitating a site that has sustained
environmental degradation,
• such as those by natural cause (desertification) and
those caused by human activity (strip mining),
• to restore that area back to its natural state as a
wildlife home and balanced habitat.
 What are some ways types of land restoration?

• Land rehabilitation.
• Environmental restoration.
• Reforestation, Forest restoration, Forest landscape
restoration.
• Restoration ecology.
• Floodplain restoration.
• Riparian zone restoration.
• Stream restoration.
• Daylighting (streams
 Chapter 7
Application of microbe in medical
biotechnology
 Sources of antibiotics and other secondary
metabolites
• Microorganisms
– Bacteria
– Fungi
• Plants
 Chapter 7

• Application of microbe in medical

biotechnology
Secondary metabolites from plants
• Research in the area of plant tissue culture technology
has resulted in the production of many pharmaceutical
substances for new therapeutics.
• Advances in the area of cell cultures for the production
of medicinal compounds has made possible the
production of a wide variety of pharmaceuticals like
alkaloids, terpenoids, steroids, saponins, phenolics,
flavanoids, and amino acids.
• Successful attempts to produce some of these valuable
pharmaceuticals in relatively large quantities by cell
cultures are illustrated.
 Secondary metabolites from microorganisms

• Microbial metabolites can be primary or

secondary metabolites
1. Primary metabolites= secreted during
active growth or exponential phase of the
organisms such as amino acids, vitamins,
organic acids, sugars, lipids, nucleic acids,
and their derivatives essential for the basic
metabolic activities. Hence, these are called
growth dependent metabolites
29
2. Secondary metabolites =are produced by slow
growing or non-growing cells of microorganisms
during depletion of one or more nutrients in the
culture medium
• They have no play role in growth of microorganisms,
and they are not further required by the
microorganisms for their growth.

• Therefore, secondary metabolites are also termed as

growth independent.

Some examples of microbial secondary metabolites

includes; antibiotics, toxins, alkaloides, etc.
30
 Antibiotics
• Antibiotics are chemicals produced by a
microorganism that kills or inhibits the growth of
another desired microorganism. But not kill the cells
in human body.
• Therefore, antibiotics are products of one organism
that kill another organism.
• Antibiotics are antimicrobial agents: i.e. they are
chemical that kills or inhibits the growth of
microorganisms
• Each different type of antibiotic affects different
bacteria in different ways.

31
 Source of antibiotics

• Bacteria= e.g. Actinomycetes, specially genus

streptomyces=
major sources
• Fungi= e.g. Aspergillaceae
• Plants = e.g. Garlic

32
 History of Antibiotics

Alexander Fleming

• Fleming was a Scottish scientist who

discovered penicillin.
• He grew colonies of Staphylococcus
aeureus in a Petri dish. A spec of mold
fell on the dish, and over time began to
grow more and more
• Fleming noted that the more the mold
grew, the levels of S. aeureus decreased.
• From this Fleming noted that the mold
must secret a fluid that had the ability to
kill the bacteria.
• The specific name of the mold was
Penicillin notatum, and thus Flemming
called the liquid secreted, penicillin.
Alexander Fleming
 Zone of Inhibition
• Around the fungal colony
is a clear zone where no
bacteria are growing
• Zone of inhibition due to
the diffusion of a
substance with antibiotic
properties from the
fungus
 Applications of antibiotics

• Particularly important as anti microbial agents

Other applications of antibiotics
• Anti-tumer, e.g. Actinomycin D
• Food preservation, e.g. Nisin
• For control of plant diseases, e.g. Tetranactin,
blasticidin
• Used in animal feed and veterinary medicines, e.g.
Tylocin, Hygromycin B
• Used as tools in molecular biology, i.e. some of the
antibiotics can selectively inhibit certain biological
reactions at the molecular level. Therefore, certain
antibiotics have been used to obtain information on
DNA replication 36
Modes of Antimicrobial Action
 Antibiotic Resistance
Why micro-organisms developing resistance?
• Unrestricted and often inappropriate use of most
antibiotics is the main reason why antibiotics are
rapidly losing their efficiency
• Antibiotic resistance can be also occur by a mutation
of DNA in bacteria or DNA acquired from another
bacteria that is drug-resistant through transformation.
Penicillin-resistant bacteria can alter their cell walls
so penicillin can not attach to it.
• etc.

38
 The Need for New Antibiotics
Why the need for new antibiotics?\
• for more effective treatment of the diseases
• to produce antibiotics with wider antimicrobial
spectrum
• to reduce toxicity of the antibiotics
• to produce antibiotics with low allergic reactions
• to decrease antibiotic resistance of
microorganisms
 Why the need for new antibiotics?...

• The problem of multiple resistance to existing

antibiotics
• The development of previously non-pathogenic
microorganisms into pathogens
• Need to develop anti-fungal antibiotics
• Need to develop antibiotics specifically for
agricultural purposes
• Need for anti-tumor and anti-parasitic drugs
 Genetic Engineering for developing of new
antibiotics
• In this process scientists are able to take
control of the biosynthetic machinery of
certain bacteria and use this information
• to create antibiotics
• to destroy harmful bacteria

41
 Strain improvement
• Several options are open to an industrial microbiology
organization seeking to maximize its profits in the face of its
competitors’ race for the same market. The organization may
undertake more aggressive marketing tactics, including more
attractive packaging while leaving its technical procedures
unchanged. It may use its human resources more efficiently and
hence reduce costs, or it may adopt a more efficient extraction
system for obtaining the material from the fermentation broth.
The operations in the fermentor may also be improved by its
use of a more productive medium, better environmental
conditions, better engineering control of the fermentor
processes, or it may genetically improve the productivity of
the microbial strain it is using.
• Of all the above options, strain improvement appears to be the
one single factor with the greatest potential for contributing to
greater profitability.
Targets or the need for industrial strain improvement

A. Increase product concentration (i.e. the metabolite

concentrations produced by wild strains are too low for
commercial processes)
B. Process improvement:
• decrease fermentation time
• be able to metabolize inexpensive substrates
• do not produce undesirable by products e.g. pigments or
substance chemically related to the main product
• reduce oxygen needs
• etc.
C. New product development: Changes in the genotype of
microorganisms can lead to the biosynthesis of new metabolites.

43
 Methods of Improvement for Industrial Strains

• Strain improvement for any application can be

performed by:
1. Traditional or classical methods. E.g.
 Select the strain from existing populations
(i.e. naturally occurring variants which over-
produce the desired product are sought.)
 Mutation using chemicals or radiation (i.e.
manipulation of the existing genetic apparatus
in a producing organism)

44
2. Genetic Engineering (rDNA techniques) (i.e.
introducing new genetic properties into the organism)
• This is the most significant approaches to
strain improvement. Because it has the
potential for not only increasing yields but also
for producing entirely new substances.
Vaccines
• are preparations of dead or weakened (attenuated)
pathogenic microorganisms, or their parts (fractions of
them) or their products that can be given to humans or
animals to produce antibodies against a specific disease
causing organisms.
• are an antigenic preparation that stimulates the immune
system to synthesize antibodies to protect the body against
infection without causing the disease.
• the antigenic material present in a vaccine is normally an
inactivated form of the infectious agent.
• Hence, vaccines are immunizing agents that trigger the
body's immune system to produce antibodies against a
specific disease causing organisms (viruses, bacteria or
other parasite).
Vaccines can be:

i. Non recombinant (traditional) vaccines

ii. Recombinant (modern) vaccines

47
i. Traditional Vaccines
• are commonly made from intact pathogenic organisms
or incompletely purified products of such organisms.
• vaccines produced by without any manipulation of
genes.
 Traditionally prepared vaccines includes:
• Attenuated live vaccine
• Killed vaccines,
• Bacterial toxoids
• Subunit or cellular fraction (polysaccharide and
polypeptide) vaccines
• Live attenuated (avirulent) vaccines- Virulent pathogenic
organisms are treated to become attenuated and avirulent but
antigenic. They have lost their capacity to induce disease but
retain their immunogenicity.
• Inactivated (killed) vaccines- when organisms are killed or
inactivated by heat or chemicals but remain antigenic. They
are usually safe but less effective than live attenuated
vaccines.
• Toxoids- They are prepared by detoxifying the exotoxins of
some bacteria rendering them antigenic but not pathogenic.
The antibodies produces in the body as a consequence of
toxoid administration neutralize the toxic moiety produced
during infection rather than act upon the organism itself. In
general toxoids are highly efficacious and safe immunizing
agents.
• Subunit or cellular fraction (polysaccharide and
polypeptide) vaccines- contain purified antigens rather
than whole organisms. i.e. they are prepared from extracted
cellular fractions.
e.g. meningococcal vaccine from the polysaccharide
antigen of the cell wall, the pneumococcal vaccine
from the polysaccharide contained in the capsule of
the organism, and hepatitis B polypeptide vaccine.
The traditional production of vaccines has
several limitations (drawbacks)

• Cell cultures are costly to maintained

• The yield of vaccines is very low
• There is the danger of non-virulent organisms (atteneuated)
being converted to virulent ones. Vaccinization by such
organisms may cause the disease itself.
• It is not possible to develop vaccines for the organisms not
grown in culture
• It is not possible to prevent all the disease by use of
traditional vaccines. E.g. AIDS

51
Q. How we can overcome the limitation of
traditional vaccine production?
Ans. By applying rDNA technology
• Before the advent of recombinant DNA technology,
vaccines were made exclusively from infectious
agents that had been either killed or attenuated. Both
types of vaccines were potentially dangerous because
they could be contaminated with the live, infectious
agent. In fact, in a small number of instances, disease
has actually been caused by vaccination. By
recombinant DNA techniques, these antigenic proteins
can be produced, completely free of the infectious
agent, and used in a vaccine.
52
ii. Recombinant vaccines
• They are vaccines produced by manipulation of genes
(Recombinant DNA technology).
Types of recombinant vaccines
• Subunit recombinant vaccines- these are the components of the
pathogenic organisms. They include proteins, peptides, and
DNA.
• Atteneuated recombinant vaccines- these are the genetically
modified pathogenic organisms (bacteria or viruses) that are
made non-pathogenic and used as vaccine. Attenuated vaccines
often consists of a pathogenic strains in which the virulent genes
are deleted or modified but organism is still able to stimulate an
immune response.
• Vector recombinant vaccines - these are genetically modified
viral vectors that can be used as vaccines against certain
pathogens.
Biotechnology has produced new and powerful vaccine.

Recombinant virus has the

outside coat of the harmful
virus, when injected into
body, the immune system
produces antibodies against
the coat of recombinant
virus.
Recombinant-Vector Vaccines

in 1986 -the recombinant vaccine

against Hepatitis B Virus (HBV) is
approved.
in 2006 - a recombinant vaccine
against human papilloma virus
(HPV) receives FDA approval. The
virus causes genital warts and can
cause cervical cancer.
DNA vaccine
• DNA sequence used as a vaccine.
• This DNA Sequence code for antigenic protein
of pathogen.
• As this DNA inserted into cells it is translated to
form antigenic protein. As this protein is foreign
to cells , so immune response raised against this
protein. In this way, DNA vaccine provide
immunity against that pathogen.
 DNA vaccines are the newest vaccines and are
still experimental. i.e. There are no DNA
vaccines in market at present. 56
Recombinant vaccines in transgenic plants
• The advent of pharming has led to the possibility of using
transgenic plants as the hosts for synthesis of
recombinant vaccines.

Edible vaccines
• Transgenic plants engineered for vaccine production can be
used as edible vaccines if the vaccines are directly
expressed in the edible parts of the plant. If the plants are
transformed with the gene responsible for vaccine
production attached with the organ-specific promoter, it
will express, and the raw, edible part can be eaten for
vaccination. The vaccine molecules present in the raw food
(tuber, fruit, or leaves) will be absorbed into the cell lining
of the mouth and food canal while it reaches the stomach. 57
Edible vaccines: minimize need for expensive refrigeration,
distribution and administration techniques.
Q. Compare and contrast production of
pharmaceuticals by traditional and modern (rDNA
technology) method
• Write the limitations of traditional method
Q. List some advantages as you can think of for
producing transgenic plants containing edible
vaccines.
Ans. Some possible advantages are:
1. the vaccines can be easily delivered without requiring local
experts
2. the vaccines do not need to be shipped and stored: plants
expressing the vaccines could be grown locally
3. there may be less resistance to eat edible vaccine than to having
an injection (some religions specifically forbid injection)
4. production could be very cheap.
B. rDNA technology in Diagnosis of diseases

Generally, disease can be diagnosed by either;

i. Traditional diagnostic method (indirect and not
specific method) - used to diagnose:
 parasite infection- using microscopic examination, in
vitro culture technique or detection of antibodies in
serum.
Drawback- time taken to culture microorganisms, difficult
to grow viruses and some bacteria in culture medium.
 genetic disease- by estimation of metabolites
(blood/urine) and by enzyme assay.
Drawback- diagnosis is carried out after appearance of
symptoms
 Chapter 8

Environmental Biotechnology
Environmental biotechnology

• is the solving of environmental problems through the

application of biotechnology.
• Is the development, use and regulation of biological systems
for remediation of contaminated environments (land, air,
water), and for environment-friendly processes.
• Environmental biotechnology applies the metabolism of
animals, microorganisms, and plants as a means of cleaning
up polluted air, soil, and water by using a strategy called
bioremediation.
• is the multidisciplinary integration of sciences and
engineering in order to utilize the huge biochemical
potential of microorganisms, plants and parts thereof for the
restoration and preservation of the environment and for
the sustainable use of resources. 62
 Why environmental biotechnology?

• It is needed to:
 eliminate the hazardous wastes (pollutants)
produced by our other technologies.
 create alternative energy sources (i.e. Bioenergy).

63
• microbe-based environmental clean up=
bioremediation.
• Certain microorganisms, including bacteria,
fungi, and algae, and sometimes higher
organisms such as plants, can use compounds
that we consider pollutants as a food source.
• Thus, they consume these compounds and
excrete less harmful substances.
Biostimulation
• is the process by which enhancement of microbial
activity by increased supply of nutrients or by
addition of certain stimulating agents such as electron
acceptor, surfactants, etc during biodegradation or
bioremediation.
Bioaugmentation
• refers to the increment (increase) of biodegradation
through manipulation of genes (i.e. use of selected
natural microorganisms isolated from the
environment and genetically engineered to improve
the process of bioremediation).

65
What is the difference between Biostimulation
and Bioaugmentation?

Biostimulation involves the modification of the environment

to stimulate existing microorganisms capable of bioremediation.
Because indigenous populations may not be capable of degrading
the xenobiotics or the wide range of potential substrates present in
complex pollutant mixtures.

Bioaugmentation is the introduction of a selected group of

natural microbial strains or a genetically engineered variant to treat
contaminated soil or water.

66
Bioremediation
• Any process that uses microorganisms including
Bacteria, fungi, green plants or their enzymes to clean
up contaminated soil or water by break down of
pollutants in order to return the environment to its
original natural condition.

67
• The use of plants to clean up the environment, known
as phytoremediation, is also considered a type of
bioremediation.
• Therefore, the use of living green plants for the
removal of contaminants and metals from soil is
known as phytoremediation.
• Terrestrial, aquatic and wetland plants and algae can
be used for the phytoremediation process under
specific cases and conditions of hydrocarbon
contamination.
• e.g. Sunflowers can removed Cesium and Strontium.

68
Phytoremediation
A conceptual view of a phytoremediation system, with a cut-away
section of the root-soil zone. When organic matter (OM) is released
from the plant roots, co-metabolic processes can be carried out more
efficiently by microbes, leading to enhanced degradation of
contaminants. The degradation of hexachlorobenzene is shown as an
example. 69
 Types of bioremediation

1. In situ bioremediation- involves a direct

approach for the microbial degradation of
xenobiotics at the site of pollution.
2. Ex situ bioremediation - the waste or toxic
materials can be collected from the polluted sites
and the biodegradation with appropriate
microorganisms can be carried out at designed
places.

70
Pollution
• is the introduction of contaminants into an
environment ( e.g. water, soil, air) that causes
instability, disorder, harm or discomfort to the
ecosystem i.e. physical systems or living organisms.

Pollutant
 is a waste material that pollutes air, water or soil.
 Three factors determine the severity of a pollutant:
• its chemical nature
• the concentration, and
• the persistence
 Pollutants
Any waste material that pollutes air, water or
soil.
 Pesticides
 Industrial Smoke Stacks
 Automobile Emissions
 Deliberate Discharge of
Compounds into Water

72
Sewage
• is the liquid waste or waste arising mainly from domestic
(residential, institutional, commercial) and industrial sources.
The most important components of sewage are:
i. Organic compounds- e.g. carbohydrates, proteins, lipids,
amino acids, urea, etc.
ii. Inorganic compounds- e.g. suspended (floatable) particles
iii. Living organisms- e.g. pathogenic organisms
 Therefore, the process designed for the treatment of sewage
should have the following objectives:
a. Treatment of biodegradable organic materials
b. Removal of floatable and suspended particles
c. Eliminations of disease causing (pathogenic) organisms.
73
 Types of water based on their human use
There are 2 kinds of water:
1. Drinking water
2. Waste water
Drinking water or potable water is water of sufficiently high
quality that can be consumed or used without risk of immediate
or long term harm. It is provided by water supply networks or
may be found in deep wells or springs.
Waste water or sewage comprises liquid waste discharged by
domestic residences, commercial properties, industry, and/or
agriculture and can encompass a wide range of potential
contaminants and concentrations. In the most common usage, it
refers to the municipal wastewater that contains a broad spectrum
of contaminants resulting from the mixing of waste waters from
different sources.
Water pollution
• Water pollution is defined as any physical, biological,
or chemical change in water quality that adversely
affects living organisms.
• Water pollution can occur three ways
– Physically
– Chemically
– Biologically
• There are many different potential sources of water
pollution, but commonly all sources are divided into
two distinctive types:
1. Point source pollution
2. Non-point source pollution
75
1. Point source pollution: refers to contaminants that
enter a waterway through a discrete conveyance, such
as a pipe or ditch. Examples of sources in this
category include discharges from a sewage treatment
plant, a factory, or a city storm drain.
2. Non-point source pollution: refers to diffuse
contamination that does not originate from a single
discrete source. NPS pollution is often a cumulative
effect of small amounts of contaminants gathered
from a large area. Nutrient runoff in storm water from
"sheet flow" over an agricultural field or a forest.
 Source of Water Pollution

1. Point source pollution - 2. Nonpoint source pollution -

source is from drain pipes, runoff from farm fields and
ditches, sewer outfalls, feedlots, lawns and gardens, golf
factories and power plants - courses, construction sites,
easy to monitor and atmospheric deposits - no specific
regulate location so harder to monitor
and regulate
Contaminants in water may include:
1. microorganisms such as viruses and bacteria
2. inorganic contaminants such as salts and metals
3. pesticides and herbicides
4. organic chemical contaminants from
industrial processes and petroleum use
5. radioactive contaminants.
6. etc
Wastewater Treatment
• It is the process by which removal of
contaminants such as organic materials,
pathogenic organisms, and suspended particles
(inorganic compounds) from wastewater.

79
Objective of wastewater treatment

Waste water treatment has dual purpose:

1. Protection of the environment
2. Safeguarding of the human health

80
Waste water treatment systems

1. Septic Tanks system= typically treat small volumes of

waste (e.g., from a single household, small
commercial/industrial)
2. Wastewater Treatment Plants (WWTPs)= typically
treat larger volumes of municipal or industrial waste.
 The commonly used laboratory methods for the
measurement of organic matter in sewage or
wastewater are:

1. Biological oxygen demand (BOD)= is the amount of

dissolved oxygen consumed by microorganisms
during the biochemical oxidation of organic and
inorganic matter to carbon dioxide at 5-day BOD
test, at 20oC
2. Chemical oxygen demand (COD)= the amount of
oxygen necessary to oxidize the organic carbon
completely to carbon dioxide, water, and NH4+
3. Total organic carbon (TOC)-used when the
concentration of organic matter is very low
4. Theoretical oxygen demand (Th OD)
82
 Sewage or Wastewater Treatment process

Stages of wastewater treatment process

1. Primary Treatment (Physical Process)

- the removal of any large objects and non-degradable
materials
- bar screen and grit chamber techniques are employed
-wastewater flow is slowed down and suspended solids
settle to the bottom by gravity
- the material that settles is called sludge or biosolids
- Sludge from the primary sedimentation tanks is pumped to
the sludge thickener.
83
Bar Screen
- catches large objects that have gotten into sewer
system such as bricks, bottles, pieces of wood, etc.

84
• Primary treatment

Grit Chamber - removes small

rocks, broken glass, diapers,
protects pumps combs, towels, plastic bags,
and equipment syringes, etc.
from damage
Bar Screen Grit Chamber Primary clarifiers

Primary
Settling
Tank

To land Anaerobic sludge

application digestor sludge
85
2. Secondary Treatment (Microbial Process)
– Effluent from primary treatment
transferred into trickling bed, or aeration
tank
– Biological degradation of dissolved organic compounds by
using bacteria and algae to metabolize organic matter in the
wastewater
– Any excess microbes are removed and sent to solids
treatment

86
Secondary Treatment…
– Supernatant or primary effluent contains
high levels of dissolved organic load
(Biological Oxygen Demand)
– Aeration to stimulate aerobic degradation
• activated sludge reactor bacteria degrade organic
• trickling filter reactor carbon to CO2
 Secondary Treatment
• Remaining suspended solids are decomposed and
number of pathogens are reduced
Final
settling tank
Primary or clarifier
settling
tank
Aeration tank
or
Trickling filter

sludge To anaerobic
sludge digester

Sludge
digester
Gravity thickener plant Land application
1% 6% solids content
3. Tertiary Treatment (Advanced process) and
(Physico-chemical Process)
-Process used when water is to be used for
irrigation, recreation, drinking water
• Involves
– Filtration
• Very effective in removing Crytosporidium and Giardia
• 90% removal of enteric bacteria and viruses
– Coagulation (iron and aluminum salts, pH>11)
• 99% removal of enteric viruses
– Activated carbon adsorption.
– Additional disinfection
89
Tertiary Treatment (Physico-chemical Process)…
• Effluent from secondary treatment is usually
disinfected (chlorinated) before release into nearby
waterway.
• Removal of plant nutrients (nitrates and
phosphates) from secondary effluent by
chemicals, or natural/constructed wetlands.

90
 wastewater treatment…

• Primary treatment (Physical removal of large debris)

– Separation of large debris following sedimentation
• Gravel, sand, leaves
• Secondary Treatment (Microbiological conversion of
organic-C to CO2 and H2O)
-Remaining suspended solids are decomposed and
number of pathogens are reduced
• Tertiary treatment
- Involves a series of steps to further reduce organic
concentration, turbidity, N, P, metals, and pathogens
91