TRENDS IN BIOENGINEERING (QUALITATIVE)

• Muscular and Skeletal Systems as scaffolds, scaffolds and tissue engineering.

• Bio-printing techniques and materials, 3D printing of ear, bone and skin. 3D printed foods.
• Electrical tongue and electrical nose in food science, DNA origami and Biocomputing, Bio-imaging
and Artificial Intelligence for disease diagnosis.
• Bio-concrete and Bioremediation and Bio-mining via microbial surface adsorption (removal of heavy
metals like Lead, Cadmium, Mercury, Arsenic).

Muscular Systems as Scaffolds

 The muscular system in the human body comprises various types of muscles, including skeletal,
cardiac, and smooth muscles.
 Skeletal muscles play a crucial role as natural scaffolds by providing structure, support, and
movement.
 The scaffold provides a framework for the cells to grow and differentiate into new heart tissue, which
can help to repair the damaged or diseased tissue.
 One example of using muscular systems as scaffolds is in the treatment of damaged or diseased heart
tissue.

Architecture

 The entire muscle is surrounded by epimysium.

 Epimysium contains layer of connective tissue called the perimysium.
 Perimysium contains bundles muscle fibers called fascicles. Each muscle fiber is surrounded by
endomysium.
 Inside the muscle fibers, there are tiny structures called myofibrils.
 Myofibrils are made up of smaller units called sarcomeres.
  Sarcomeres contain thin (Actin) and thick filaments (Myosin) that work together to make the muscle
fibers contract.

Muscle Cells as Scaffold

By removing the living cells from the muscle tissue, leaving behind the structure known as the extracellular
matrix (ECM), can provides a framework can support the growth of new tissues.
1. Harvesting muscle tissue: Small sample of muscle tissue is taken, typically from a donor.
2. Cell removal: Living cells within the muscle tissue are removed using a process called
decellularization. Done by treating the tissue with specific chemical solutions/enzymes that break
down and wash away the cellular components, while preserving the ECM.
3. ECM scaffold: Remaining ECM is now a scaffold. It consists of proteins, such as collagen and elastin.
4. Seeding cells: Decellularized muscle scaffold is then seeded with desired cells. These can be stem cells
or specialized cells. The cells are introduced onto the scaffold, allowing them to attach and populate
the structure.
5. Growth: Over time, the seeded cells differentiate, meaning they multiply and transform into specific
cell types required for the desired tissue.
6. Tissue integration: As the cells continue to grow, they populate the scaffold and form new tissue. The
new tissue integrates with the surrounding native tissue, gradually replacing the decellularized scaffold
with functional, regenerated tissue.
Muscular Dystrophy
 It is a group of genetic disorders that result in
progressive weakness and degeneration of the skeletal
muscles, which are responsible for movement.
 The disorders are caused by mutations in genes that
encode proteins needed for muscle function.
 The most common type of muscular dystrophy is
Duchenne muscular dystrophy,Becker muscular
dystrophy, limb-girdle muscular dystrophy, and
facioscapulohumeral dystrophy, among others.

1. Duchenne muscular dystrophy (DMD)- appears in ages 2-3. Affects boys but can affect girls in rare
cases.
 Primary symptom is muscle weakness, begins in the muscles close to the body and later affects muscles
in the outer limbs.
2. Becker muscular dystrophy- appears in ages of 5-15.
 Similar to DMD, except that it progresses slower and symptoms begin to appear later, muscle loss
begins in pelvic area, thighs, and shoulders.
3. Facioscapulohumeral dystrophy (FSHD) typically appears before the age of 20 in both males and
females.
 FSHD affects the muscles of the face, around the shoulder blades, and in the upper arms.
 Currently no cure for muscular dystrophy, but various treatments can help manage symptoms & slow
progression of disease.

Bioengineering solutions for muscular dystrophy

Some of the approaches being explored include:
1. Gene therapy: Delivering a functional copy of missing or mutated gene to affected muscle cells. Goal
is to restore the production of missing protein and improve muscle function.
2. Stem cell therapy: Using stem cells to replace damaged muscle cells and promote repair and
regeneration of the muscle tissue.
 Stem cells can be taken from the patient's own body (autologous stem cells) or from a donor (allogenic
stem cells).
3. Exoskeleton technology: Using wearable devices, such as robotic exoskeletons, to support & enhance
movement of individuals with muscular dystrophy.
 Devices use motors and sensors to mimic the movements of wearer & help improve mobility.
4. Tissue engineering: Using a combination of materials (scaffolds & growth factors) to promote growth
& repair of muscle tissue.
  Goal is to create functional muscle tissue to replace damaged tissue in individuals with muscular
dystrophy.

Skeletal System
 It refers to framework of bones, joints, and connective
tissues that provide structure, support, and protection
to the body.
The key components and functions of the skeletal system are:
 Bones: Human body consists of 206 bones that vary in
size and shape. Composed of hard and dense
connective tissue that provides strength and support.
 They serve as the anchor points for muscles, protect
internal organs, and store minerals like calcium and
phosphorus.
 Cartilage: Flexible connective tissue found in certain joints and structures such as the ears and nose.
 Acts as a cushion between bones, reducing friction & absorbing shock.
 Ligaments: Tough bands of fibrous tissue that connect bones to other bones in joints, providing
stability and preventing excessive movement.
 Tendons: Tendons are strong fibrous tissues that connect muscles to bones, enabling movement by
transmitting the force generated by muscles.
Axial Skeleton: The axial skeleton forms the central axis of the body and includes the skull, vertebral column,
and ribcage.
 The skull protects the brain, and the vertebral column (spine) supports the body's weight and houses
the spinal cord. The ribcage encloses and protects the heart, lungs, and other thoracic organs.
Appendicular Skeleton: Comprises bones of the limbs and shoulder & pelvic girdles.
 Upper limbs (arms) consist of upper arm bone, forearm bones and the hand bones. The lower limbs
(legs) include thigh bone, lower leg bones, and the foot bones.
 The shoulder and pelvic girdles attach the limbs to the axial skeleton.

Skeletal System as Scaffold

 The skeletal system can be used as a scaffold for tissue growth in certain applications.
 Natural structure of the skeletal system can serve as a scaffold or template for tissue engineering
purposes. Synthetic/natural scaffolds, designed to mimic properties of bone, can be used to fill void
left by a bone defect.
 Scaffold provides a 3D structure support for attachment, proliferation, & differentiation of cells
involved in bone regeneration.
 Over time, scaffold can be replaced by newly formed bone tissue, resulting in restoration of bone
structure & function.
Osteoporosis

 Condition that weakens bones & makes them more likely to break (fracture), especially the bones in
the hip, spine, and wrist.
 It occurs when body loses bone mass and density more quickly than it can be replaced, leading to
fragile bones that are prone to fracture.
 Common condition, especially among older women, and it can increase the risk of falls and fractures.
 Risk factors for osteoporosis include older female, having a family history of the condition, smoking,
drinking excessive amounts of alcohol, being thin or having a small body frame, and having a low
calcium intake.
Bioengineering solutions for osteoporosis
Some of the approaches being explored include:
1. Tissue engineering: Involves using scaffolds and growth factors to stimulate the growth of new bone
tissue and promote the repair of damaged bones.
 The goal is to create functional bone tissue that can replace the lost bone mass and density.
2. Stem cell therapy: Using stem cells to replace damaged bone cells and promote the repair &
regeneration of bone tissue.
 Stem cells can be autologous stem cells or allogenic.
3. Iomaterials: Using synthetic/natural materials to replace or augment damaged bone tissue.
Biomaterials can be designed to mimic the properties of natural bone and promote the growth of new
bone tissue.
4. Gene therapy: Delivering a functional copy of a gene involved in bone growth and repair to the
affected bone cells. Goal is to restore the production of the missing protein and improve bone health.

Bioprinting Techniques and Materials

Introduction
  Bio-printing is a rapidly growing field that uses various techniques to produce three dimensional (3D)
structures and functional biological tissues for medical and scientific applications.
 The main objective of bio-printing is to mimic the structure and function of human tissues and organs,
leading to the development of replacement parts for damaged or diseased organs.

Comparison between 3D Printer and Bio-printer

Bio-printing Materials
 Bio-printing materials, also known as bio-inks, are specifically designed to be compatible with
living cells and provide a supportive environment for their growth and organization.
 Here are some examples of commonly used bio-printing materials:
 1. Hydrogels: Hydrogels are water-based polymer networks that closely mimic the extracellular matrix
(ECM) found in living tissues.
2. Cell-laden Aggregates: In some cases, cells are first aggregated into biomolecules and biomaterials (or
microtissues) before being incorporated into the bio-ink.
3. Decellularized Extracellular Matrix (dECM): The extracellular matrix (ECM) is a complex network of
molecules surrounding cells in tissues and organs.
4. Synthetic Polymer-based Bio-inks: Synthetic polymers can be used to create bio-inks with well-
defined mechanical properties and degradation rates.
5. Composite Bio-inks: Composite bio-inks combine different materials to enhance the bio-ink's
properties, such as mechanical strength, printability, and cell behavior.
Most Commonly used Bio-printing Techniques
1. Inkjet-based Bio-printing: works similarly to standard inkjet printing.
2. Extrusion-based Bio-printing: uses a syringe or a similar mechanism to extrude the bio-ink through
a nozzle.
3. Laser-assisted Bio-printing: utilizes laser energy to precisely deposit bio-inks onto a substrate.
4. Microvalve-based Bio-printing: employs microvalves to control the deposition of bio-inks.
5. Bio-printing with Solid Freeform Fabrication: combines bio-printing with traditional 3D printing
methods.

3D Printing of Ear
 Using a 3D printer to create an ear-shaped structure with a special material, like biocompatible
polymer/hydrogel, as the "ink."
 The printed ear structure is then seeded with human cartilage cells, which grow and develop into
functional ear tissue over time.
 Main advantage of is it allows for creation of an ear that is custom-fitted to an individual patient,
based on their specific ear shape and size.
Materials Used for 3D Printing of Human Ear
 Some of the most commonly used materials for 3D printing of ears include:
 1. Hydrogels: Soft, gel-like materials, commonly used in bioprinting due to their ability to mimic
mechanical properties of human tissues.
2. Biocompatible polymers: Synthetic materials that are compatible with human tissues and do not cause
adverse reactions, ex: Polylactide (PLA).
3. Scaffolds: Structures that provide a supportive framework for the cells to grow and develop.
4. Cell-embedded materials: Materials that contain living cells, which can be used to seed the 3D
printed structure.
5. Ceramics: Such as hydroxyapatite, can be used in 3D printing of ear structures.
Technological Importance
1. Personalized ear prosthesis: 3D printing allows for the creation of customized ear prostheses that
match the unique anatomy of each patient.
2. Faster production and lower costs: Traditional methods of ear prosthesis fabrication can be time-
consuming and expensive.
3. Biocompatibility: 3D printing can use biocompatible materials for the production of ear prostheses,
reducing the risk of adverse reactions and improving patient outcomes.
4. Medical education: 3D printing of human ears can be used to educate medical students and
healthcare professionals on the anatomy and treatment of ear defects and injuries.
3D Printing of Bone
 This process involves using a 3D printer to create a bone-shaped structure using a special material,
such as a biocompatible polymer or a ceramic material, as the "ink."
 The printed bone structure can then be implanted into a patient to replace missing or damaged bone
tissue.
 There are two main approaches to 3D printing of bones:
1. Additive manufacturing: Additive manufacturing involves building up the bone structure layer
by layer. The layer-by-layer deposition of material enables the creation of complex three-
dimensional structures that mimic the natural bone tissue.
2. Scaffold-based techniques: creating a porous structure that provides a framework for bone cells
to grow and develop. These techniques involve the fabrication of biocompatible and
biodegradable scaffolds using 3D printing technology,
Materials Used for 3D Printing of Bone
Some of the most commonly used materials for 3D printing of bones include:
1. Biocompatible polymers: Synthetic materials that are compatible with human tissues and do not
cause adverse reactions. They can be used as the "ink" in 3D printing, providing a supportive structure
for the cells to grow and develop into functional bone tissue. Ex: polyethylene, polycaprolactone.
2. Ceramics: Such as hydroxyapatite, are natural components of human bones and can be used as
the "ink" in 3D printing. Ex: Hydroxyapatite, Calcium phosphate, Tricalcium phosphate.
3. Scaffolds: Structures that provide a supportive framework for the cells to grow and develop. Ex:
Polyglycolic acid (PGA), Poly-L-lactic acid (PLLA).
 4. Cell-embedded materials: Materials that contain living cells, which can be used to seed the 3D
printed structure. The cells then grow and develop into functional bone tissue over time. Ex: Gelatine
methacryloyl, Alginate.

3D Printing of Skin
 The goal of 3D printing skin is to create functional, living tissue that can be used for a variety of
purposes, such as cosmetic testing, wound healing, and drug development.
 The process involves the use of bio-printing technology, where a bio-ink made from living cells and
growth factors is printed in a specific pattern to create the desired tissue structure.

Materials used for 3D printing of Skin

1. Hydrogels: Such as alginate and collagen, are hydrophilic materials that can be used to create 3D
structures for cell growth.
2. Polymers: Such as polyethylene glycol and polycaprolactone, can be used in 3D printing of skin.
3. Cell-laden hydrogels: Materials that contain living cells and can be used to create 3D printed skin
structures.
4. Scaffolds: Structures that provide a supportive framework for cells to grow and develop.

Technological Importance
 Better wound healing: 3D printing of skin can produce customized skin grafts that promote wound
healing and reduce the risk of infection.
 Reduced scarring: 3D printed skin can promote more natural healing & reduce scarring, improving
cosmetic appearance of the skin after injury.
 Replication of skin structure: 3D printing can replicate the structure & properties of natural skin,
such as the thickness and elasticity of different layers of the skin.
 Reduced donor site morbidity: promotes faster healing.
  Alternative to animal testing: Can provide an alternative to animal testing in the cosmetic and
pharmaceutical industries, reducing the ethical concerns and improving the accuracy.
 Research and development: Can be used in research and development to study properties and
behavior of different skin types.

3D Printed Foods
 3D printed food refers to food items that are created using 3D printing technology.
 This technology allows for the creation of food items with intricate shapes and designs, which can
be customized based on individual preferences and dietary needs.
 The process of 3D printing food involves the use of edible materials, such as pastes, gels, and
powders, which are combined and printed layer by layer to create the final product.
 The use of 3D printing in the food industry has the potential to revolutionize the way food is produced,
as it allows for the precise control of portion sizes and ingredients, which can be beneficial for
individuals with specific dietary needs or restrictions.
 Additionally, it can be used to create unique and customized food items that would be difficult to
achieve using traditional cooking methods.

Materials used for 3D Printing of Food

 Edible pastes: Edible pastes, such as pureed fruit, chocolate, and cream cheese, can be used in 3D
printing of food.
 Edible gels: such as agar and gelatin, can be used in 3D printing of food. These are flexible to create
aesthetically pleasing.
 Edible powders: such as flour and sugar. Can be combined with liquids to form a printable mixture.
 Sweet and savory snacks: such as crackers, cookies, and chips, can be customized to include
intricate shapes and designs.
 Pastries: 3D printing technology can be used to create intricate and aesthetically pleasing pastries,
such as cakes and cupcakes.
 Decorative garnishes: 3D printing technology can be used to create unique and attractive garnishes
for dishes, such as cheese and fruit designs.

The importance of 3D printing in the food industry

Here are some key reasons why 3D printing is important in the food industry:
 Customization and Personalization: 3D printing enables the creation of customized and
personalized food products. It allows for the precise control of ingredients, textures, flavors, and
nutritional content, catering to individual preferences, dietary restrictions, and specific
nutritional needs.
 Novelty and Creativity: 3D printing in the food industry allows for the creation of intricate and
visually appealing food designs that are difficult to achieve with traditional food preparation methods.
 It offers the opportunity to experiment with shapes, structures, colors, and patterns, thereby
enhancing the dining experience and presentation of food.
 Enhanced Food Safety: With 3D printing, the entire food production process can be tightly
controlled and automated, reducing the risk of contamination and human error. The technology
allows for the use of fresh, high-quality ingredients and eliminates the need for excessive processing
and preservatives.
 Supply Chain Efficiency: 3D printing has the potential to streamline the food supply chain by
enabling on-demand production. It eliminates the need for long-distance transportation and storage
of certain food products, reducing food waste and improving overall efficiency.
 Sustainable Food Production: 3D printing has the potential to reduce food waste by using precise
ingredient measurements and optimizing production processes.

Electrical Tongue (E-Tongue) in Food Science

 Device used in food science to analyze the
taste and flavor of food and beverages.
 Works by measuring electrical conductivity,
impedance, & capacitance of a food/beverage
sample, which are related to the concentration
of ions in the sample and the texture of the
sample.
 This technology allows for rapid & non-
invasive analysis of food/beverages, as it does
not require human taste testers.
 The e-tongue provides a numerical
representation of taste & flavor of sample, which can be used to compare and analyze different food
and beverage products.
Working & Components of E-Tongue
Consists of a sensor array, which is placed in contact with the food/beverage sample.
1. Sensor Array- Refers to a collection of multiple sensors that are designed to detect and measure
different taste qualities. These sensors are often specific to particular taste components and provide
information about the presence and intensity of specific taste attributes.
2. Sample holder- Holds the food sample during testing. Made of inert material which does not interact
with food sample (such as glass or plastic).
3. Signal processor- Responsible for analyzing electrical signals generated by sensor array in response
to the food sample. It converts these signals into a profile of the taste and flavor of the sample.
4. Data analysis software- Interpret data generated by signal processor & create a profile of the taste &
flavor of food sample.
5. Calibration solutions- Ensure accurate & reliable results e-tongue requires regular calibration using
standardized solutions with known taste and flavor characteristics.
 E-Nose in Food Science
 Known as an electronic/electrical nose, is a technology used in food science for the analysis and
characterization of food and beverage aromas and flavors.
 The electrical nose typically consists of a sensor array that is capable of detecting and quantifying
volatile organic compounds (VOCs) in food and beverage.
1. Sensor Array- The e-nose sensor array is composed of several sensors that are sensitive to specific
VOCs each sensor is designed to respond to a particular chemical or group of chemicals and together
they can provide a comprehensive profile of the Aroma are order.
2. Sample delivery system- Used to introduce sample into the e-nose.
3. Signal processing system- Analyzes electrical signals generated by sensor in response to the sample.
4. Data analysis software- Interpret data by the signal processing system and create the profile of the
aroma/odor of the sample.
5. Calibration solutions- Ensure accurate & reliable results e-nose requires regular calibration using
standardized solutions with known taste and flavor characteristics.
DNA Origami & Bio-computing
 DNA Origami is a technique in nanotechnology that involves folding DNA molecules into specific
shapes. The process involves using a long, single strand of DNA, called the scaffold, to guide the
folding of short, complementary DNA strands, called staples, into a desired shape.
 Bio-computing refers to the use of biological systems, such as cells, enzymes, and DNA, for computing
and information processing. This field combines the principles of computer science, biology, and
engineering to create novel systems for computing and data storage.
 Templates for organizing and positioning of biomolecules can be designed by DNA origami structures
which allows for the creation of DNA “nanobots” which performs functions like sensing, delivery
drugs to specific location of the body and performing computations.
 One of the major advantages of DNA origami in bio computer is the ability to program the interactions
between different biomolecule in precise manner.
 For example DNA Nano robots created by this technique can detect & destroy cancer cells by
delivering the drug to the location.
 DNA origami also used to create molecular logic gates, which are basic building blocks of digital
computing. DNA based computing has the potential to be faster, smaller, & more energy efficient
than the Silicon based computing.
 DNA origami can be used as a tool for biomedical imaging, as it can be designed to target specific
cells or tissues and provide high-resolution images.
 Bio-computing systems can be used to monitor and track environmental conditions, such as air and
water quality, in real-time.
 Bio-computing systems can be used to store and process large amounts of data, as DNA has a high
information density [consider that a single gram of DNA can theoretically store up to 215 petabytes (1
petabyte = 1 million gigabytes) of data].
 Bio-computing systems are energy-efficient, highly robust, as they are less susceptible to errors and
failures compared to traditional electronic systems
Bio-imaging for Disease Diagnosis
 Bio-imaging is the use of imaging technologies to visualize biological processes and structures in
living organisms.
 It plays a crucial role in disease diagnosis by providing detailed images of the body's internal structures
and functions, and can help healthcare professionals to identify and diagnose a wide range of diseases
and conditions.
 Some examples of bio imaging techniques used for disease diagnosis include X-rays, CT scans, MRI,
PET scans, ultrasound, and optical imaging. These technologies can be used to visualize a wide range
of structures and functions, including bones, tissues, organs, blood vessels, and more.
 Bio-imaging can be used to detect diseases in their early stages, when they are often more treatable.
This can lead to earlier treatment and better outcomes for patients.
 Bio-imaging technologies can be combined to provide a multi-modal view of the body's internal
structures and functions, which can provide a more comprehensive understanding of a disease or
condition.

Artificial Intelligence for Disease Diagnosis

 Artificial Intelligence (AI) has the potential to revolutionize the field of disease diagnosis by providing
healthcare professionals with more accurate and efficient tools for identifying and treating various
conditions.
 AI algorithms can analyze medical images, such as X-rays, CT scans, and MRIs, to detect signs of
diseases.
 For example, AI algorithms can identify patterns in medical images that may indicate the presence of
a particular condition, such as a lung cancer, breast cancer or heart disease. This type of image analysis
is known as computer-aided diagnosis (CAD).
 AI algorithms can analyze a patient's medical history, lifestyle habits, & genetic information to
recommend the best course of treatment for their condition.
 AI algorithms can be integrated into electronic health records to provide physicians with information
about the best diagnostic tests & help with real-time decision-making.
 Google help achieved level of accuracy comparable to an experience radiologist in detecting breast
cancer from mammography images and the efficiency of detection is 89%.
 AI algorithms can identify early signs of diabetic retinopathy allowing doctors to intervene before the
disease progresses.
 In combination with bio imaging AI has the potential to revolutionize disease diagnosis.

Bio mineralization
  It is a process by which living organisms form minerals or mineralized structures within their tissues.
 Process is carried out by specialized (mineralizing) cells which form & organize minerals in specific
shapes sizes and functions.
 Mineralizing cells produce organic matrices (templates for mineralization).
 Matrices are composed of proteins, lipids, & other bio molecules which can interact with minerals to
control their growth.
 Mineral ions are transported to the mineralizing cells either by surrounding environment or within
organisms body.
 Mineral ions are then deposited on to the matrix & begins crystalline structure.
 These crystalline structures continue to grow and organize. Ex: seashells, bones, teeth & coral reefs.

Self-Healing Bio-concrete
 Self-healing bio-concrete is a type of
concrete that incorporates
microorganisms, such as Bacillus
fragments, into the mixture, along
with calcium lactate as a nutrient
source.
 The microorganisms are activated
when the concrete cracks, and they
produce calcium carbonate, which
fills in the cracks and repairs the
concrete.
 Bio-concrete include increased
durability, reduced maintenance
costs, and improved sustainability,
as the concrete is able to repair
itself without the need for human
intervention.
 Additionally, because the microorganisms used in the concrete are naturally occurring and non-toxic,
self-healing bio-concrete is considered to be environmentally friendly.
 When the concrete cracks, water and oxygen enter the crack and activate the Bacillus bacteria.
 The bacteria then produce calcium carbonate, which is a type of mineral that is commonly found in
natural stone.
 The calcium carbonate acts as a binder and fills in the cracks, repairing the concrete and restoring its
structural integrity.
 The Bacillus bacteria are also able to survive in a wide range of temperatures and pH levels, making
them well-suited for use in concrete.

Bioremediation and Bio mining via Microbial Surface Adsorption

 (Removal of heavy metals like Lead, Cadmium, Mercury, Arsenic)
 Bioremediation and bio mining are two related but distinct processes that utilize living organisms to
clean up contaminated environments or extract valuable minerals, respectively.
 Bioremediation refers to the use of microorganisms, plants, or animals to clean up contaminated
environments, such as soil, water, or air. This process occurs naturally over time, but can also be
accelerated through the addition of specific microorganisms or other biotic agents.
 Bio mining, on the other hand, refers to the use of microorganisms to extract valuable minerals from
ore deposits. This process involves the use of microorganisms to dissolve minerals from ore, creating
a solution that can be separated and purified to obtain the valuable minerals.
 Bioremediation and bio mining via microbial surface adsorption is a process that utilizes
microorganisms to remove heavy metals like lead, cadmium, mercury, and arsenic from contaminated
environments or ore deposits, respectively.
 Microbial surface adsorption is a method that utilizes microorganisms to adsorb and immobilize heavy
metals from contaminated solutions. This process involves the use of microbial cells, like bacteria or
algae which have affinity from metal ions, allowing them to adsorb and sequester these metals on their
cell surfaces.

Process of removing polluting heavy metals

1. Identification of heavy metal-contaminated site:
• Identify the site or area contaminated with heavy metals, such as soil, water, or industrial waste sites.
2. Isolation and characterization of metal-resistant microbial strains:
• Select and isolate microbial strains that have demonstrated resistance to heavy metals. These can
include bacteria / fungi.
3. Culturing and enrichment of microbial strains:
• Culture and propagate the selected microbial strains in a suitable growth medium under laboratory
conditions. This step aims to obtain a sufficient quantity of active microbial biomass for subsequent
applications.
4. Preparation of microbial suspension:
• Harvest the microbial biomass and prepare a suspension (serve as the delivery system) by suspending
the biomass in a carrier solution, such as water.
5. Application of microbial suspension to the contaminated site:
• Apply the microbial suspension to the heavy metal-contaminated area. This can be done through
spraying, injection, or soil/water mixing, depending on the specific site conditions.
6. Microbial adsorption and sequestration of metal:
• Through their metabolic activity, the microbes produce extracellular compounds such as organic acids
or biofilm matrix components that have an affinity for binding metal ions.
7. Separation or removal of metals from the contaminated site can be achieved through different
methods
Lead
Examples of Microbes Used:
 Pseudomonas sp.: Some strains of Pseudomonas bacteria have the ability to tolerate and accumulate
lead.
 Bacillus sp.: Certain Bacillus species have been found to exhibit resistance to lead and can effectively
bind and remove it.
Cadmium
Examples of Microbes Used:
 Cupriavidus metallidurans: This bacterium is known for its high resistance to heavy metals, including
cadmium.
 Trichoderma sp.: Some species of Trichoderma fungi have shown the ability to tolerate and accumulate
cadmium.
Mercury
Examples of Microbes Used:
 Pseudomonas putida: Certain strains of Pseudomonas putida have the ability to tolerate and accumulate
mercury.
 Penicillium chrysogenum: Some strains of Penicillium chrysogenum fungi have shown the capacity to
bind and remove mercury.
Arsenic
Examples of Microbes Used:
 Shewanella sp.: Certain strains of Shewanella bacteria have the ability to tolerate and accumulate
arsenic.
 Aspergillus niger: Some strains of Aspergillus niger fungi have shown the capacity to bind and remove
arsenic.