CHAPTER 1

INTRODUCTION

1.1 GENERAL
Water is one of the most important substances on earth. All plants and
animals must have water to survive. If there was no water there would be no life
on earth1 .It covers 71% of the
Earth’s surface, and is vital for all known forms of life. But only 2.5% of
the Earth’s water is fresh water.Rapid urbanization and industrialization releases
enormous volumes of wastewater, which is increasingly utilized as a valuable
resource for irrigation in urban and peri-urban agriculture This growing problem
of water scarcity has significant negative influence on economic development,
human livelihoods, and environmental quality throughout the world. So it has
become anessential need for today’s environment to protect water from getting
polluted or develop its cost effective remedial method for its protection. The
various conventional methods for waste water treatment are present since the
ancient time4-10 but they are very costly and not economical. Therefore new
technologies are being introduced to overcome the conventional methods of waste
water treatment. At this point comes the potential of using microalgae for waste
water treatment. It has self cleansing power due to which it abstracts Nitrate 99%,
sulphate 84% and Phosphate 73% for its growth and development. During their

growth they trap sun light and CO2 from the environment for their photosynthesis.
It is also been found that wastewater treatment using microalgae has number of
positive applications over the conventional methods as it is useful in Wastewater

treatment, CO2 sequestration, Cost effective, Sanitation and also in the production
of renewable sources of energy such as Biodiesel, Biofuel, Glycerol, Methane gas,
Hydrogen gas, Biofertilizersetc.The use of microalge for treatment of waste water
and its advantages are discussed in this report.

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 CHAPTER 2

WASTEWATER CHARACTERISTICS

2.1 GENERAL
Wastewater can be characterized by its physical, chemical and biological
component of wastewater.Physical, chemical and biological methods are used
today to remove contaminants from wastewater. The main goal of wastewater
treatment management is the protection of the environment and the public health
by protecting the water bodies which provide important water resources. Table 1
shows the composition of untreated domestic wastewater levels.
2.1.1 Physical characteristic

 Solids : Solids are classified into three main types: 1. Total Solids (TS): All
the matter that remains as residue upon evaporation at 103oC to 105oC. 2.
Settleable solids: Settleable solids are measured as ml/L, which is an
approximate measure of the sludge that can be removed by primary
sedimentation. 3. Suspended solids (SS) and Filterable solids (FS).

 Odour :Odour is produced by gas production due to the decomposition of

organic matter or by substances added to the wastewater.

 Temperature :Temperature of wastewater is commonly higher than that of

water supply. Depending on the geographic location the mean annual
temperature varies in the range of 10 to 21oC with an average of 16 oC.

 Density:-Almost the same density of water when the wastewater doesn't

include significant amount of industrial waste.

 Color: Fresh waste water-light brownish gray.

 With time- dark gray
 More time- black (septic).
 Some times pink due to algae or due to industrial colors
 Turbidity :It's a measure of the light – transmitting properties of water.

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2.1.2 Chemical characteristic
Points of concern regarding the chemical characteristics of wastewater are:
 Organic matter
 Measurements of organic matter
 Inorganic matter
 Gases
 pH

2.1.2.1 Organic matter (cahboc )

Organic mater is derived from animals & plants and man activities.
 75% SS - organic. (Suspended Solids)
 40% FS-organic. (Filtered Solids)

 Proteins (40-60%).
 Carbohydrates (25-50%).
 Fats, Oils, and Grease (10%).

2.1.2.2 Inorganic matter

The main inorganic materials of concern in wastewater treatment are:
 Chlorides
 Nitrogen
 Phosphorous
 Sulphur
 Toxic inorganic compounds (copper,lead,silver,chromium,arsenic,boron)

 Heavy metals
(nickels,manganese,lead,chromium,cadmium,zinc,copper,iron,
mercury.

2.1.2.3 Gases
The main gases of concern in wastewater treatment are
nitrogen,oxygen,carbon dioxide, hydrogen sulphide, ammonia, methane.

2.1.2.4 pH

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 The hydrogen-ion concentration is an important parameter in both natural
waters and wastewaters. It is a very important factor in the biological and
chemical wastewater treatment

2.1.3 Biological characteristic

Wastewater naturally holds large amount of macro and microorganism.
Determining biological treatment depends on the quantity of any species of macro
and micro organism and aquatic life in a receiving water bodies. Within treatment
facilities, wastewater provides or aerobic wastewater treatment.The main
microorganisms of concern in wastewater treatment are Bacteria, Fungi, Algae,
Protozoa, Viruses, and pathogenic microorganisms groups.

2.2 SEWAGE TREATMENT PROCESSES

2.2.1 Preliminary treatment of sewage

The preliminrary treatment of sewage removes large solid materials by
sewers that could flow through the plant or damage equipment . these materials
are composed of floating equipments such as rags,wood,fecal materials and
heavier grit particles. Large floating objects can be removed by passing the
sewage through 20-60 spaced , the retained materials is raked from the bars at
regular intervals . Grit is removed by reducing the flow velocity to a range at
which grit and slit will settle, but leave organic material in suspension, this is
usually in the velocity range of 0.2-0.4 m/s.

2.2.2 Primary treatment of sewage

After removal of coarse materials, sewage passes to sedimentation tank,
which aim to remove the settleable solids(represent upto 70% of the total
settleable soilds) by gravity.a well designed sedimentation tank can remove 40%
of BOD in the form of settleable soilds.

2.2.3 Secondary treatment of sewage

The secondary treatment aims to reduse the BOD exerted by reducing

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organic matter. This is mediated, primarily, by a mixed population of heterotrophic
bacteria that utilize the organic constituent of energy and growth.
A large number of biological unit operations are available to achieve the
aerobic oxidation of BOD.several investigators have pointed out that biological
oxidation systems can remove over 90% of pathogenic bacteria from sewage,
however the removal of viruses is much more variable.

2.2.4 Tertiary treatment of sewage

Tertiary treatment process aims to remove all organic ions. it can
be accomplished biologically or Chemically. the biological treatment process
appears to perform well when compared to chemical processes which are in
general too costly to be implemented in most places an which may lead to
secondary pollution. in addition, each additional treatment step in a waste water
system greately increases the total cost

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 CHAPTER 3

WASTEWATER TREATMENT USING MICROALGAE

3.1 GENERAL
The concept of wastewater treatment using microalgae was firstly
proposed by Oswald in the 1950, and the concept was later expanded to propose
use of this system for energy production through harvesting and utilization of
algal biomass. Shirai F, et. al. in the year 1998 who investigated the treatment of
Soy Sauce effluent including four types of microalgae in which they applied this
method for the fermentation of ethanol from the biomass which was obtained by
harvesting microalgae. After that so many scientist turned their research work to
treat various industrial and municipal waste water using different types of
microalgae. Bio-treatment with microalgae is particularly attractive because of
their photosynthetic capabilities, converting solar energy into useful biomasses
and incorporating nutrients such as nitrogen and phosphorus causing
eutrophication . This fascinating idea launched some fifty-five years ago in the
U.S. It has since been intensively tested in many countries .
3.2 MICROALGAE FOR WASTEWATER TREATMENT
The land-space requirements of microalgal wastewater treatment systems
are substantial , efforts are being made to develop wastewater treatment systems
based on the use of hyperconcentrated algal cultures. This proved to be highly
efficient in removing N and P within very short periods of times, e.g. less than 1
h .The algal systems can treat human sewage , livestock wastes , agro-industrial
wastes and industrial wastes. Also, microalgal systems for the treatment of other
wastes such as piggery effluent , the effluent from food processing factories and
other agricultural wastes have been studied. Also, algae based system for the
removal of toxic minerals such as lead, cadmium, mercury, scandium, tin, arsenic
and bromine are also being developed .The technology and biotechnology of
microalgal mass culture have been much discussed . Algal systems have
traditionally been employed as a tertiary process. They have been proposed as a
potential secondary treatment system .Tertiary treatment process removes all
organic ions. It can be accomplished biologically or chemically. The biological

6
tertiary treatment appears to perform well compared to the chemical processes
which are in general too costly to be implemented in most places and which may
lead to secondary pollution. However, each additional treatment step in a
wastewater system greatly increases the total cost. The relative cost of treatment
doubles for each additional step following primary treatment .A complete tertiary
process aimed at removing ammonia, nitrate and phosphate will thus be about four
times more expensive than primary treatment. Microalgal cultures offer an elegant
solution to tertiary and quinary treatments due to the ability of microalgae to use
inorganic nitrogen and phosphorus for their growth . And also, their capacity to
remove heavy metals , as well as some toxic organic compounds , therefore, does
not lead to secondary pollution. Amongst beneficial characteristics they produce
oxygen, have a disinfecting effect due to increase in pH during photosynthesis.
Algae can be used in wastewater treatment for a range of purposes, some of which
are used for the removal of coliform bacteria, reduction of both chemical and
biochemical oxygen demand, removal of N and/or P, and also for the removal of
heavy metals.

Figure 2.1 : Simple schematic representation of wastewater treatment

using microalgae and obtained byproduct

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3.3 PRINCIPLE OF MICROALGAE BASED WASTEWATER
TREATMENT
The main pollutants in a variety of wastewater sources are Nitrogen (N)
and Phosphorus (P) in different forms, which contribute to eutrophication of
bodies of water in lakes, rivers or oceans worldwide if discharged improperly. On
the other hand, these pollutants are ideal nutrients to stimulate fast algae growth.
The typical metabolic processes in microalgae include mechanisms of light
harvesting, carbon acquisition through photosynthesis, and N and P assimilation
as well as formation of unique secondary metabolites (e.g., DHA and EPA, unique
pigments and active compounds, etc.) for different applications. It is worth noting
that although most microalgae in nature are obligate photoautotrophs and their
growth strictly depends on photosynthetically derived energy, there are some
species that can grow in a Heterotrophic Mode (HM) in the presence of organic
carbon under complete darkness or in a Mixotrophic Culture Mode (MM) when
supplied with both organic and inorganic carbon under light/dark conditions .
Thus the main mechanisms for nutrients removal in wastewater by microalgae
include typical photosynthetic assimilation and/or chemosynthetic assimilation by
heterotrophic/mixotrophic metabolic pathway as well as abiotic factors such as
ammonia volatilization and phosphorus precipitation through elevated pH.

3.4 MECHANISMS OF MICROALGAE IN WASTEWATER TREATMENT

Many characteristics of microalgae make them very attractive for
wastewater treatment. Apart from removing nutrients, microalgae have been
shown to remove coliforms and heavy metals, and reducing BOD.
The nitrogen removal capabilities of microalgae is based on a process
called assimilation. In this process, inorganic forms of nitrogen are converted to
their organic forms . Assimilation is performed by all eukaryotic algae, at the level
of the cell membrane. As all inorganic nitrogen forms are converted to ammonia
prior to incorporation into amino acids, ammonia seems to be the preferred form
of nitrogen. This has also been shown experimentally. Ammonium can also be
passively removed by ammonium stripping. In ammonium stripping, ammonia is
volatized by the elevated pH due to microalgal photosynthesis and/or elevated

8
temperature.
- 2-
The preferred forms of phosphorus for uptake are H 2PO4 and HPO4 .
These forms are actively transferred across the plasma membrane and
incorporated into organic compounds through phosphorylation (e.g. to create
ATP). Some algae have also been shown to be able to use phosphorus from
organic esters for growth. Similar to nitrogen, phosphorus can also be removed
passively by the increased pH. At these increased pH levels, phosphates have the
tendency to precipitate with multivalent cations.

Coliform bacteria are rod-shaped, Gram-negative, non-sporing and motile

bacteria, who can ferment lactose at body temperature (37°c). They are often used
as a bacterial indicator for water quality, and are usually found in the faeces of
warm-blooded animals. While they themselves are normally not problematic, their
presence often indicates the presence of pathogens, such as other bacteria, viruses
and protozoa. These coliforms, and thus also their pathogenic colleagues, are very
often found in wastewater effluents.
Scenedesmusobliquuscultures could totally remove coliforms such as E.
coli from domesticsewage in four days. The inhibitory effect on coliforms was
attributed mainly to elevation of pH. Also it showed reduction of faecal bacteria in
the presence of microalgae.
Another interesting feature of microalgae in light of water treatment is
their capability of removing heavy metals. Heavy metals can be removed by
microalgae in a process called metal sorption. In this process, the heavy metals
bind either to the cell surface or to intracellular ligands, and can thus be removed
from the water by harvesting the algal biomass. Heavy metals that can be removed
this way are zinc, chromium, cadmium, cobalt, aluminium, copper and mercury.
Also removal of other contaminants, such as herbicides and fungicides, has been
shown.
Lastly, hypoxia in natural water bodies is mainly created by the
metabolising activity of heterotrophic bacteria. The amount of organic carbon in
the water can be expressed as COD or BOD (chemical and biological oxygen
demand respectively). These indicate how much oxygen is needed to chemically

9
or biologically oxidize the available carbon molecules. Excess BOD often leads to
high metabolic bacterial activity and rapid depletion of oxygen levels. The role of
microalgae in reducing BOD levels lies in their photosynthetic capacities. During
photosynthesis oxygen is produced, thus preventing hypoxia in the water.

3.5 NUTRIENT REMOVAL CAPABILITY OF MICROALGAE

A major requirement in wastewater treatment is the removal of nutrients
and toxic metals to acceptable limits prior to discharge and reuse. Algae are
autotrophs, i.e. they can synthesize organic molecules themselves from inorganic
nutrients. A stoichiometric formula for the most common elements in an average

algal cell is C106H181O45N16P, and the elements should be present in these

proportions in the medium for optimal growth. Microalgae have been proven to be
efficient in removing nitrogen, phosphorus, and toxic metals from a wide variety
of wastewaters. There are extensive studies of algae growth in municipal
agricultural and industrial wastewaters. Substantial amounts of nutrient removal
and algae biomass production were obtained in these studies. Hence, controlled
microalgae cultivation shows promise as a potential biological treatment method
for wastewater.This integrated wastewater treatment and biofuel production
system can thus benefit the community as well as the environment.
Although, nitrogen and phosphorous are the two main nutrients of concern
in eutrophication, being limiting factors in most growth scenarios, other
micronutrients, including silicon and iron, can affect the abundance of
phytoplankton communities. How- ever, many of the micronutrients are toxic to
most algae species at high concentrations. Some of them also form precipitates
with other essential elements and reduce their availability. However some algae
strains are particularly tolerant to heavy metals and their potential to absorb metals
has been demonstrated

3.5.1 Nitrogen removal

Nitrogen is a critical nutrient required in the growth of all organisms.
Organic nitrogen is found in a variety of biological substances, such as peptides,
proteins, enzymes, chlorophylls, energy transfer molecules (ADP, ATP), and

10
genetic materials (RNA, DNA). Organic nitrogen is derived from inorganic

sources including nitrate (NO3), nitrite (NO2), nitric acid (HNO3), ammonium

(NH4), ammonia (NH3), and nitrogen gas (N2). Micro- algae play a key role in
converting inorganic nitrogen to its organic form through a process called
assimilation. In addition, cyanobacteria are capable of converting atmospheric
nitrogen into ammonia by means of fixation.
3.5.2 Phosphorous removal
Phosphorus is also a key factor in the energy metabolism of algae and is
found in nucleic acids, lipids, proteins, and the intermediates of carbohydrate
metabolism. Inorganic phosphates play a significant role in algae cell growth and
metabolism. Phosphates are transferred by energized transport across the plasma
membrane of the algal cell. Not only are inorganic forms of phosphorus utilized
by microalgae, but some varieties of algae are able to use the phosphorus found in
organic esters for growth.

Figure 2.2 : Microalgae wastewater treatment releases effluent after treatment

containing very low nitrogen and phosphorous

3.6 LOW ENERGY REQUIREMENT

Traditional wastewater treatment processes involve the high energy costs
of mechanical aeration to provide oxygen to aerobic bacteria to consume the
organic compounds in the wastewater, whereas in algae based wastewater

11
treatment, algae provides the oxygen for aerobic bacteria. Aeration is an energy
intensive process, accounting for 45 to 75% of a wastewater treatment plant’s total
energy costs. Algae provide an efficient way to consume nutrients and provide the
aerobic bacteria with the needed oxygen through photosynthesis. Roughly one kg
of BOD removed in an activated sludge process requires one kWh of electricity

for aeration, which produces one kg of fossil CO2 from power generation . By
contrast, one kg of BOD removed by photosynthetic oxygenation requires no
energy inputs and produces enough algal biomass to generate methane that can
produce one kWh of electric power.

3.7 COST EFFECTIVENESS

It has been shown to be a more cost effective way to remove biochemical
oxygen demand, pathogens, phosphorus and nitrogen than activated sludge
process and other secondary treatment processes. Cost for using other chemicals
for removal of organic ions can be avoided. Similarly cost for a separate unit for
aeration can also be saved since algae gives out oxygen through photosynthesis.

3.8 PRODUCTION OF USEFUL ALGAL BIOMASS

The resulting algae biomass is a source of useful products such as
biodiesel. Previous research in the early 1990’s by the National Renewable Energy
Laboratory (NREL) showed that under controlled conditions algae are capable of
producing 40 times the amount of oil for biodiesel per unit area of land, compared
to terrestrial oilseed crops such as soy and canola .However, their results also
showed that large-scale algae cultivation for energy production was uneconomical
at that time and suggested future research into waste - stream integration . It is
hoped that the economics will be ultimately improved by combining biodiesel
feedstock production with agricultural or municipal wastewater treatment and
CO2 fixation.

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 Figure 2.3 : Useful algal biomass obtained after wastewater treatment
using microalgae

(source: Google images)

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3.9 THE GREEN HOUSE GASES EMISSION REDUCTION
Carbon, in the form of carbon dioxide, may be fixed from the atmosphere
and industrial exhaust gases through the photosynthetic activity of autotrophic
microalgae. Carbon can be also utilized in the form of soluble carbonates for cell
growth, either by direct uptake or conversion of carbonate to free carbon dioxide
through carbo anhydrase activity.
The use of algae to mitigate carbon dioxide from flue gases is another
research focus and, if effective, could benefit both the environment and biofuel
production.The Us Environmental Protection Agency (EPA) has specifically
identified conventional wastewater treatment plants as major contributors to
greenhouse gases. Algae based waste water treatment also releases CO2 but the
algae consume more CO2 while growing than that is being released by the plant,
this makes the entire system carbon negative.

-----

Figure 2.4 : CO2and sunlight absorbed by microalgae in

wastewater treatment

(source: Google images)

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3.10 REDUCTION IN SLUDGE FORMATION
In conventional wastewater treatment systems the main aim is to minimize
or eliminate the sludge. Industrial effluents are conventionally treated using a
variety of hazardous chemicals for pH correction, sludge removal, colour removal
and odour removal. Extensive use of chemicals for effluent treatment results in
huge amounts of sludge which forms the so called hazardous solid waste
generated by the industry and finally disposed by depositing them in landfills. In
algae wastewater treatment facilities, the resulting sludge with algal biomass is
energy rich which can be further processed to make biofuel or other valuable
products such as fertilizers. Algal technology avoids use of chemicals and the
whole process of effluent treatment is simplified. There is considerable reduction
in sludge formation

3.11 REMOVES HEAVY METALS

Another interesting feature of microalgae in light of water treatment is
their capability of removing heavy metals. Heavy metals can be removed by
microalgae in a process called metal sorption. In this process, the heavy metals
bind either to the cell surface or to intracellular ligands, and can thus be removed
from the water by harvesting the algal biomass. Heavy metals that can be removed
this way are zinc, chromium, cadmium, cobalt, aluminium, copper and mecury.
Also removal of other contaminants, such as herbicides and fungicides, has been
shown.

Algae can be used to make bioethanol and biobutanol and by some estimates can
produce vastly superior amounts of vegetable oil, compared to terrestrial crops
grown for the same purpose. Algae can be grown to produce Hydrogen. In 1942 a
German researcher named Hans Gaffron, while working at the University of
Chicago, observed that the algae he was studying, Chlamydomonasreinhardtii (a
green algae), would sometimes switch from the production of Oxygen to the
production of Hydrogen. Algae can be grown to produce biomass, which can be
burned to produce heat and electricity.

15
3.13 OTHER APPLICATIONS

Algae can be used to make bioethanol and biobutanol and by some

estimates can produce vastly superior amounts of vegetable oil, compared to
terrestrial crops grown for the same purpose. Algae can be grown to produce
Hydrogen. In 1942 a German researcher named Hans Gaffron, while working at
the University of Chicago, observed that the algae he was studying,
Chlamydomonasrenhardtii ( a green algae), would sometimes switch from the
production of Oxygen to the production of hydrogen. Algae can be grown to
produce biomass, which can be burnt to produce heat and electricity.

3.14 FACTORS AFFECTING ALGAL GROWTH AND NUTRIENT

REMOVAL
Algal growth and nutrient uptake are not only affected by the
availability of nutrients, they also depends on complex interactions among
physical factors such as pH, light intensity, temperature and biotic factors. The
first abiotic factor significally influencing algal growth is the light intensity, it is
expected that the higher the algal density, better the growth and higher the nutrient
removal efficiency. However the high algae content would leadto self shading an
accumulation of autoinhibitors and a reduction in photosynthesis efficiency .

16
 CHAPTER 4

CASE STUDIES

4.1 AQUACULTURE INDUSTRY

KENT BIOENERGY FACULTY NEAR MECCA, CA

The company now holds a variety of patents and exclusive licenses for
aguaculture wastewater treatment systems, algae-based water recycling systems,
and algae-based environment remediation technology. It also has patents pending
for making algae easier to harvest, methods for maintaining algae monocultures
( ensuring that a pond has just one species of algae), and for genetically modifying
algae to enhance algal production of valuable oils that can be used to make fuels.

4.2 CHEMICAL INDUSTRY

CHEMFAB ALKALIS Ltd. PONDICHERY, INDIA
The factory uses ground water during processing at various stages to extract useful
chemicals from crude sea salt. The effluent water that is discouraged has a very
high salinity and TDS. A team carried out research work on treatment of waste
water employing various species of micro algae including a few freshwater as well
as marine forms to remove nutrients and bring down TDS. Successful tesults were
obtained with immobilized cell of micro algae. The work is underway to extend
this technique to field conditions.

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 CHAPTER 4

CONCLUSIONS
3.1 GENERAL

Microalgae have the natural wastewater treatment properties. It has the self
cleansing power due to which it abstracts Nitrate around 90%, sulphate 80% and
Phosphate 70% for its growth and development. During their growth they trap sun
light and CO2 from the environment for their photosynthesis. In the mean time of
review of literature we had found that waste water treatment using microalgae has
number of positive applications over the conventional methods as it is useful in
Wastewater treatment, CO2 sequestration, Cost effective, Sanitation and also in
the production of renewable sources of energy such as Biodiesel, Biofuel,
Glycerol, Methane gas, Hydrogen gas, Biofertilizers etc.

4.2 CONCLUSION

The overall review has concluded that this Green technical method for
treatment of municipal waste water using microalgae should be applied in all
developing and developed countries for wastewater treatment so as to protect the
environmental pollution causing due to waste water from industrial and domestic
effluents. It is also seen that the biomass obtained from these treatment plants can
be changed to useful by-products like bio fuels, animal feeds etc, hence it is
profitable and also a renewable energy source.

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REFERENCES
 I. Rawat, R.Ranjith Kumar, T.Mutanda, F.bux, 2010, Dual role of micro
algae: phycomediation of domestic wastewater and Biomass production for
sustainable Biofuels production, Elsevier
 N.Abdel-Raouf, A.A.Al- himaidan, I.B.M.Ibrahim, 2012, Microalgae and
wastewater treatment, Saudi journal of biological science
 S.R.Lyon,H.ahmedzadeh and M.A. Murry, 2015, Algae-based wastewater
treatment for biofuel production: Processes, species and extraction methods.,
springer international publishing Switzerland
 S.N.Nandeswar and G.D. Satpute, 2014, Green technical methods for
treatment of wastewater using microalgae and its application in the
management of natural water resources: A review, current world environment,

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