SEMINAR REPORT

submitted by

GOKUL NATH G S
20619026

to

Cochin University of Science and Technology

in partial fulfilment of the requirements for the award of B. Tech Degree

in Safety and Fire Engineering.

Division of Safety and Fire Engineering

School of Engineering
IMMOBILIZED MICROALGAL SYSTEM: AN ACHIEVABLE IDEA FOR
UPGRADING CURRENT MICROALGAL WASTEWATER TREATMENT
SEMINAR REPORT

submitted by
GOKUL NATH
20619026
to

Cochin University of Science and Technology

in partial fulfilment of the requirements for the award of B. Tech Degree

in Safety and Fire Engineering.

Division of Safety and Fire Engineering

School of Engineering

Cochin University of Science and Technology

2023
DIVISION OF SAFETY AND FIRE ENGINEERING
 SCHOOL OF ENGINEERING

COCHIN UNIVERSITY OF SCIENCE AND TECHNOLOGY

CERTIFICATE

Certified that this report entitled Immobilized microalgal system: An achievable idea for
upgrading current microalgal wastewater treatment is the report of seminar presented by
GOKUL NATH G S, 20619026 during 2019-2023 in partial fulfilment of the
requirements for the award of the Degree of Bachelor of Technology in Safety and Fire
Engineering of Cochin University of Science and Technology.

Mrs. Nithya Gopinath Dr. Renjith V R

Division of Safety and Fire Engineering Head

School of Engineering Division of Safety and Fire
Engineering School of Engineering
 ACKNOWLEDGEMENTS

I take this opportunity to thank the supreme being, the source of all knowledge whose
blessings are my guiding light in any venture I take up.
I am short of words to express my gratitude to Mrs. Nithya Gopinath and Mr. Praveen
Vijayan, who guided me and helped me constantly with their inputs and suggestions, to
bring out the best within me and thus the seminar report title.
I am highly indebted to Dr. Renjith V R, head of the Division for his constant supervision,
support, and motivation, for teaching me the value of integrity and hard work not only in
taking this seminar but also in any responsibility towards life.
I also express our heartfelt gratitude to my parents for being constantly motivating and
supportive.
A bouquet of gratitude to all the teachers to provide me with solutions to my problems.

GOKUL NATH G S
20619026

i
 ABSTRACT

The article discusses the urgent global issue of efficient and sustainable wastewater
treatment. One promising solution is using microalgae to treat wastewater, which can also
create a valuable biomass resource. The review proposes upgrading current microalgal
wastewater treatment by using immobilized microalgae, which involves fixing them in place
so they can be used more efficiently for treating pollutants in water. The article analyzes
various technical approaches and mechanisms involved with this process, as well as potential
applications on a larger scale. Overall, immobilization could help make microalgal-based
wastewater treatment more effective and realistic while offering new research directions for
future studies in this field. Furthermore, from high-tech technologies to promote large-scale
production and application potentials in diverse wastewater and bioreactors to downstream
applications lead upgradation closer, the feasibility of upgrading existing microalgal
wastewater treatment into immobilized microalgal systems is thoroughly discussed.
Eventually, several research directions are proposed toward the future immobilized
microalgal system for microalgal wastewater treatment upgrading. Together, it appears that
using immobilization for further upgrading the microalgae-based wastewater treatment can be
recognized as an achievable alternative to make microalgal wastewater treatment more
realistic.

ii
 CONTENTS

SL NO TOPIC PAGE

NO
1 Acknowledgment ⅰ
2 Abstract ⅱ

3 List of figures ⅳ

4 List of tables ⅴ

5 Introduction 1
6 CHAPTER 2-immobilized microalgal wastewater treatment systems 2
7 2.1. Benefits and drawbacks of immobilized microalgae 3
8 2.2. Pollutants removal by immobilized microalgae
9 2.3. Effects of immobilization on the physiological activity of microalgae 5

10 2.3.1. Enhancement of cell growth and morphology 6

11 2.3.2. Positive responses of metabolism, physiology, and productivity 7

12 2.3.3. Protection of cells against environmental stresses or toxicity 8

13 CHAPTER -3. main technical approaches employed for microalgal 9

immobilization
3.1. Adsorption or attachment onto a matrix

3.2. Entrapment within a porous matrix

iii
 LIST OF FIGURES

FIGURE PAGE NO

7.1) Survey Locations 4

iv
 LIST OF TABLES

TITLE PAGENO
8.1) Mobile Phone Distraction 5

8.2) Association between mobile phone and other risky crossing behaviour 6

8.3) Impact of mobile phone distraction on conflict situation 7

v
 CHAPTER 1-INTRODUCTION

The negative effects of water pollution are a significant concern in many countries.
According to the United Nations, over 2212 km3 of wastewater is produced annually
worldwide, which corresponds to 56% of global water withdrawals. Direct discharge of
untreated wastewater into the environment poses risks such as human health threats and
ecosystem disruption. Conventional methods for treating wastewater consume extra
chemicals and energy while emitting greenhouse gases. In China, higher effluent standards
have led to increased operating costs and chemical consumption for biological treatment
processes like activated sludge process or sewage sludge production that inevitably produces
dual attributes "pollution" and "resource." If not handled safely these can become another
source of pollution leading upgradation closer by using immobilization techniques on
microalgae-based systems seems achievable alternative method with future research
directions proposed towards it being applied more widely in diverse applications including
large-scale production facilities downstream from bioreactors where feasible references exist
already available through this review article provided herein published under Creative
Commons license CC BY-NC-ND4.

Researchers are exploring new ways to treat wastewater sustainably, and using microalgae
for photosynthetic treatment has become popular. This method is energy-efficient and
combines nutrients removal from wastewater with the advantages of microalgae. However,
this process can be affected by environmental conditions such as light intensity, which affects
carbon metabolism in microalgae. Additionally, harvesting these tiny cells from low-density
distribution within wastewater is difficult and costly. Immobilized cultivation may provide a
solution to these issues by preventing free movement of cells while retaining desired
biological activity in limited space for reuse in an aqueous phase system with high-efficiency
treatment capabilities that lower carbon footprint without extra recycling operations or
downstream risks posed by other methods like flocculation processes used during suspended
cultivation techniques currently available on large-scale production levels at low cost yet
effective pollutant removal efficiency rates remain elusive goals among researchers seeking
vi
solutions through bio-technology-based immobilization technologies over the last decade
towards sustainable biological waste-water treatments worldwide.
vi
i
 CHAPTER 2-IMMOBILIZED MICROALGAL WASTEWATER
TREATMENT SYSTEMS

The text discusses the use of immobilized microalgae for wastewater treatment.
Immobilization is inspired by natural microorganisms' ability to self-aggregate and adhere to
surfaces, which can be reproduce artificially with microalgal cells. This method has
ecological advantages similar to biofilms and can remove nutrients from wastewater while
also making it easier to harvest the cells for further use. Without immobilization, harvesting
these cells would be difficult and limit its applicability in treating wastewater using
microalgae-based methods.

2.1. Benefits and drawbacks of immobilized microalgae

Immobilized cells have a higher cell density, which allows for better removal of pollutants
such as nitrogen and phosphorus from wastewater. They also have improved light utilization,
leading to higher cell growth potential. The use of immobilization can help microalgal cells
adapt to environmental stresses or toxicity and reduce the toxicity of certain nano-adsorbents
used in treating wastewater containing nitrogen and phosphorus. Immobilized microalgae can
resist cell growth disruption, avoid photo-inhibition and reduce cellular toxicity. Bigger
immobilized beads or carriers make traditional harvesting and dewatering processes easier
and more energy-saving. Immobilization reduces the interference of introducing alien
microorganisms to the original ecosystem because it inhibits their liberation into wastewater.
However, polymers or carriers in immobilization systems may inhibit mass transfer and
resource absorption while also increasing operational costs compared to a suspended system.
Additionally, potential secondary environmental pollution risks caused by prolonged
operation might hinder its applicability for wastewater treatment purposes.

2.2. Pollutants removal by immobilized microalgae

The potential of using microalgae for wastewater treatment due to their ability to efficiently
vi
convert nutrients into value-added biomolecules.ii Immobilized microalgae may be particularly
useful in this regard because they are easy to harvest and have higher environmental
resistance. The removal rate of various pollutants by non-suspended microalgae varies
greatly, but COD, TP, and NH4+ -N tend to be removed more easily. Some types of
microalgae can also deal with other pollutants like PPCPs, plastics, heavy metals, dyes,
antibiotics and pharmaceuticals. The effects of immobilization on the physiological activity
of microalgae must be understood in order for it to become a viable option for wastewater
treatment.

FIG 1

2.3. Effects of immobilization on the physiological activity of microalgae

Immobilized microalgae can have different performance depending on the materials used for
immobilization. Some synthetic foams and resins may contain leftover pre-polymers that are
highly toxic, while other polymers may be non-toxic. The process of immobilizing or
encapsulating microalgae in polymers can also impact their growth due to chemical
interactions and limitations between the matrix and cells, as well as possible metabolite
accumulation within the matrix. Mass transfer limitations caused by the matrix thickness,
metabolic by-product accumulation, light availability, and CO2 resistance are key factors
affecting physiological changes in immobilized microalgae. However, natural non-toxic
polymers are increasingly being used for immobilization which creates a beneficial physical
barrier against diverse wastewater conditions; therefore, it is important to understand how
this affects physiological activity of microalgae when developing new methods for treating
wastewater using an immobilized system instead of suspended systems.

2.3.1. Enhancement of cell growth and morphology

ix

This section discusses how immobilizing microalgae in polymers or carriers can be beneficial
for their growth and morphology. This is important because it helps to increase the efficiency
of using microalgae for wastewater treatment. Researchers have found that entrapping cells in
hydrogels or packed bed column reactors can lead to higher growth rates and biomass yields
compared to suspension cultures, demonstrating potential applicability. Co-immobilization
with other microorganisms has also been proven as a functional strategy for increasing the
population of microalgae, leading to enhanced proliferation, pigment production, and
biomass yield. Phytohormones produced by plant-growth-promoting bacteria may play a vital
role in stimulating microalgal growth when co-immobilized together with them.

2.3.2. Positive responses of metabolism, physiology, and productivity

Liu et al. found that immobilization can alter cell growth behavior, carbon utilization, and
nitrogen uptake while maintaining stable amino acid synthesis in Chlor- ella vulgaris under
mixotrophic conditions. Calderon et al. achieved a higher biomass yield with similar cellular
components when comparing an immobilized Botryococcus braunii culture to a suspended
one, and also reported improved lipid yield and fatty acid composition compared to free cells
which may be useful for biofuel production according to Abu Sepian et al.Huang et al.'s study
showed
increased anabolic activity of microalgae as well as dye decolorization capacity through co-
immobilizing them with bacteria using calcium alginate beads.

2.3.3. Protection of cells against environmental stresses or toxicity

Immobilized microalgae can withstand environmental stresses and toxicity better than free-
floating microalgae. Immobilization positively affects the growth and physiological activity
of microalgae, but it may also limit their growth due to low mass transfer of CO2 or nutrients.
Researchers have found that stock culture management using alginate-encapsulated algal
spores is a vital part of wastewater treatment as it maintains normal physiological activities
and faster growth compared to non-encapsulated spores even under adverse conditions like
climate change, radiation, or corrosion.

x
 CHAPTER -3. MAIN TECHNICAL APPROACHES EMPLOYED FOR
MICROALGAL IMMOBILIZATION

3.1. Adsorption or attachment onto a matrix

The use of immobilization techniques to fix microalgal cells on different types of carriers for
ecological applications such as wastewater treatment and carbon capture. To achieve this, an
appropriate matrix is necessary that has satisfactory strength, higher porosity, low toxicity
and high stability under varied circumstances. Synthetic and natural carriers are used with
attachment mechanisms occurring through gravity or chemical binding. However, obstacles
like potential microalgal toxicity may affect treatment efficiency so directional design using
scaffolds associated with green adhesive coating techniques is required to overcome these
bottlenecks. method of creating a bio composite coating of cyanobacteria on flexible
polyester sheets using an external electric field. This material can be used to convert carbon
dioxide into fuels or chemicals using solar energy. Another technique involves depositing
microalgae-containing sodium alginate thin layers onto glass carriers and gelling them with
amino-functionalized silica sol to create a reinforced alginate hydrogel, which is better at
resisting high salt environments in mariculture wastewater treatment. Natural biomass-based
scaffold materials like loofah, corn cob, pine bark, sugarcane bagasse or cotton cloth pieces
are also explored as bio-matrix for establishing contact between microalgal cells and the
support structure quickly.

3.2. Entrapment within a porous matrix

Entrapping microalgal cells within a porous matrix is a common technique for immobilizing
them. This involves allowing the cells to diffuse into or cross-link with the matrix, which
must be highly hydrated and porous. Two types of matrices are used: polysaccharide gels
(such as alginate) and polymeric materials (such as gelatin). Alginate is commonly used due
to its low cost, minimal toxicity, high transparency, and stability. The process involves
adding microalgae to an aqueous solution containing monovalent alginate salts with
appropriate mixing before extruding it through a nozzle into a cation-containing solution for
polymerization or cross-linking mechanism formation.
xi Immobilized beads with entrapped
microalgal cells are obtained after stabilization; however, optimization of this system is
necessary in order to achieve higher resistance towards wastewater treatment volumes/types
by optimizing addition of microalgal beads against various wastewater treatments etc

FIG 2

FIG 3

xi
i
 CHAPTER-4 MECHANISMS OF POLLUTANTS REMOVAL BY
IMMOBILIZED MICROALGAE

In order to use immobilized microalgae for wastewater treatment, it is important to

understand how pollutants are removed. Researchers study two main perspectives: primary
elimination pathways and specific degradation processes. Extracellular polymeric substances
(EPS) released from microalgae can help absorb contaminants in the surrounding
environment and transport them into the microalgae cells. Enzymes play a critical role in
pollutant biodegradation, such as ammonium absorption capacity being positively correlated
with cellular activities of glutamate dehydrogenase (GDH) and glutamine synthetase (GS).
Researchers also explore substance transformation by analyzing metabolic by-products of
target pollutants after incubation with substrates or using techniques like HPLC/LC-HRMS to
theorize removal pathways involving hydroxylation, o-methylation, glycosylation,
dehydrogenation, and decarboxylation. A process called "assimilation" converts nitrogen into
organic compounds that support protein synthesis through photosynthesis and energy-transfer
molecules like ADP.
Immobilized microalgae can be used to remove pollutants from wastewater. The cells are
surrounded by a matrix or self-generated substances that act as glue and buffer media,
forming a concentration gradient of pollutants outside the cell wall. The polymers in this
system absorb, store, transfer and slow-release the pollutants into the microalgal cells for
removal through gradual sorption, accumulation within the cells and cellular biodegradation
similar to suspended cells. Microalgae-based treatment efficiently redirects nutrients from
wastewater for algal biomass production with metabolic routes summarized for better
understanding of nutrient uptake pathways.

xi
ii
Mechanism of waste removal by algae

FIG 4

xi
v
Intracellular and extracellular

FIG 5

x
v
 CHAPTER 5 -IMMOBILIZED MICROALGAL SYSTEM BE APPLIED
IN REAL WASTEWATER TREATMENT

There is a wide potential of using immobilized microalgae for wastewater treatment.

However, there are still challenges to achieving a cost-effective and stable large-scale system
that can effectively treat various types of real-world wastewater. The text raises questions
about how to achieve production goals and upgrade existing technologies with an
immobilized microalgal system.

5.1. High-tech technologies promote large-scale production.

New technologies for producing large amounts of microalgae in hydrogel membranes and
silk protein hydrogels using 3D-printing and robotic arms. These immobilized systems have
shown excellent viability, growth, photosynthetic activity, and pollutant removal capabilities.
These technologies could be used for industrial applications in areas such as bioremediation
and bioenergy production.

5.2. Application potentials in diverse wastewater and bioreactors

While there have been many studies on this topic, more research is needed to determine its
effectiveness in different types of wastewaters and at larger scales. Some examples are given
where immobilized microalgae were used successfully to remove pollutants from various
types of wastewaters using different reactors or treatment systems. The text also mentions
that immobilizing microalgae can lead to higher cell density and biomass, which could be
useful for downstream applications such as bioenergy production or recovery/reuse
processes. However, further investigation is needed into converting microalgal biomass while
simultaneously treating wastewater in order to make this technology more viable.

5.3. Downstream applications lead upgradation closer

This passage discusses the challenges of using xmicroalgae for energy production due to low
vi
biomass yields. However, immobilizing microalgae in wastewater can lead to higher cell
density and biomass, which is advantageous for downstream applications such as bioenergy
or bioactive compound production. Further research is needed on simultaneous wastewater
treatment and microalgal biomass conversion to make this technology more viable. Recent
advances in pretreating microalgal biomass are summarized by Bhatia et al., including
potential uses for immobilized microalgal biomass.

x
vi
 CHAP
TER 6-CONCLUSION

The use of immobilized microalgae to treat wastewater, which has been proposed as a
method for some time. However, there is limited research on scaling up and long-term
operation, which hinders its practical application. The article suggests that using immobilized
microalgae can have advantages over suspended cultivation in terms of higher cell density
and biomass for downstream applications such as bioenergy production or recovery/reuse.
Further investigation into simultaneous wastewater treatment and microalgal biomass
x more viable. It compares suspended and
conversion is needed to make this technology
vi
immobilized systems, explains how pollutants are removed by immobilized microalgae, and
explores possibilities for upgrading current wastewater treatment processes using this
technology. The text also identifies some challenges that need to be addressed in order to
improve the effectiveness of this approach, such as leakage of cells from the system over time
and a limited number of contaminants being investigated. Finally, it suggests that more
research is needed to determine whether or not immobilization is better than other methods
for cultivating microalgae in wastewater treatment processes.
Use of immobilized substrate as a barrier for cells in wastewater treatment. However, this
method can lower treatment efficiency by restricting nutrient entrance. It is important to
clarify how substances transfer and convert in these systems through experimental studies
and modeling explanations. Techno-economic assessments are needed to understand its
removal effectiveness and energy conversion potential, with targeted research on increasing
productivity while decreasing energy requirements. To make it more practical, integrating
microalgal immobilization with other biological technologies like activated sludge or
symbiotic bacteria is necessary along with intelligent farming using IoT, Big Data, and AI for
wider application in the future.

xi
x