BIO SAFETY STANDARDS

AND ETHICS (BT242AT)IV

SEMESTER B E BASKET COURSE
Dr G Vijayakumar Dr
PraveenKumar Gupta
Dr Trilok Chandran Dr A H
Manjunatha Reddy
Dr Lingayya Hiremath Dr
Narendra Kumar Sura
 Introduction
After Covid – 19, we have improved our quality of living when
compared to earlier.
Importance of hygiene
Personal healthcare
Food Habits and hygiene
Keeping these 3 as basic points we have designed the course on Bio
safety standard and ethics. As an Engineer of any branch should know
about Biosafety levels, Food safety and Bio ethics
 Unit I & II
Biohazards, Bio safety levels and Bio safety cabinets and their types.
Various parameters considered for design of Biosafety cabinets
Biosafety guidelines of Government of India, GMOs (Genetically
Modified Organisms) , LMOs (Living modified organisms)
Roles of Institutional Biosafety Committee, RCGM (Review
committee on Genetic manipulation)
GEAC (Genetic Engg. Approval Committee) for GMO applications in
food and agriculture.
Overview of National Regulations and relevant International
Agreements including Cartagena Protocol.
 Unit III
FSSAI (Food Safety and Standards Authority of India), Functions, License and
rules.
Food Hygiene: General principles of food microbiology and overview of
foodborne pathogens
sources of microorganisms in the food chain, Quality of foods, Microbial
food spoilage and Foodborne diseases
Overview of beneficial microorganisms and their role in food processing and
human nutrition, Food Analysis and Testing
General principles of food safety management systems, Hazard Analysis
Critical Control Point (HACCP).
 Unit IV & V
Food preservations, Processing, and Packaging.
Food Processing Operations, Principles, Good Manufacturing Practices
Overview of food preservation methods and their underlying principles
including novel and emerging methods/principles and novel packaging
materials
Food safety: Food Hazards, Food Additives, Food Allergens Drugs, Hormones,
and Antibiotics in
Animals. Factors That Contribute to Foodborne Illness, Consumer Lifestyles
and Demand, Food
Production and Economics, History of Food Safety, The Role of Food
Preservation in Food Safety.
Ethics: Clinical ethics, Health Policy, Research ethics, ethics on Animals.
Thank youfor any queries pl contactvijayakg@rvce.edu.in mobile
no 9886272998trilokc@rvce.edu.in mobile no 9591519849
 Biological hazards
Biological hazards, also known as biohazards, refer to biological
materials (microorganisms, plants, animals, or their byproducts) that
pose a threat to the health of living organisms, primarily that of
humans.
This can include medical waste or samples of a microorganism,
viruses, or toxins (from a biological source) that can affect human
health.

Potential biohazards can be found anywhere – at your job, in your doctor’s office, in your children’s classrooms –
and should be handled with extreme caution.
 Types of biological hazards
Viruses, such as Coronavirus (COVID-19) and Japanese encephalitis.
Toxins from biological sources.
Spores.
Fungi.
Pathogenic micro-organisms.
Bio-active substances
 What are some biohazard
Human blood and blood examples?
products. This includes items that have
been affected by blood and other body fluids or tissues that contain visible
blood.
Animal waste. Animal carcasses and body parts, or any bedding
material used by animals that are known to be infected with pathogenic
organisms.
Human body fluids. Semen, cerebrospinal fluid, pleural fluid, vaginal
secretions, pericardial fluid, amniotic fluid, saliva, and peritoneal fluid.
Microbiological wastes. Common in laboratory settings, examples of
microbiological wastes include specimen cultures, disposable culture dishes,
discarded viruses, and devices used to transfer or mix cultures.
Pathological waste. Unfixed human tissue (excluding skin), waste
biopsy materials, and anatomical parts from medical procedures or autopsies.
Sharps waste. Needles, glass slides and cover slips, scalpels, and IV
tubing that has the needle attached.
 How body is effected
Most biological hazards can cause disease in humans, from the
common cold to life-threatening diseases. They can also cause other
effects, such as poisonings, or provoke an allergic response.
Chronic health conditions are long-term conditions and diseases
lasting 3 months or longer. They may not have a cure.
Acute conditions - sometimes called poisonings - are adverse effects
from either a single dose of a substance, multiple doses given within
24 hours or an inhalation exposure of 4 hours.
Introduction to Bacteria and virus in
human body
The human body is made of about 100 trillion cells. These cells are quite complex. Most have a nucleus
and many special parts.
Bacteria are much simpler. A bacterium is made of only one cell but has no nucleus. Bacteria are small;
each is about 1/100th the size of a human cell.
Bacteria are like fish swimming in the ocean of your body. As they swim around, they eat and reproduce
rapidly. One bacterium can become millions of bacterium in just a few hours.
Viruses are completely different. A virus is a particle of DNA or RNA with a special cover over it. When a
virus comes in contact with a living cell, it attaches to it. The virus injects its DNA or RNA into the cell.
The virus DNA or RNA takes over and uses the cell to make more viruses.
Eventually the cell dies and bursts open spewing millions of new viruses into the body of its victim. Each
new virus particle can infect another cell
Bacteria
Bacteria are microscopic, single-celled organisms that can live anywhere. They are classified as prokaryotes, a
simple internal structure that lacks a nucleus. They contain DNA that either floats freely in a twisted, thread-like
mass called the nucleoid, or in separate, circular pieces called plasmids. Not all bacteria are harmful, some are
necessary for healthy body function.
Bacteria are classified according to a range of criteria including the nature of their cell walls (i.e. gram positive or
negative), their shape (i.e. round, cylindrical or spiral), or by differences in their genetic makeup.
Most bacteria multiply by a process called binary fission – a single bacterial cell makes a copy of its DNA, doubles
its cellular content, and then splits apart pushing the duplicated material out creating two identical ‘daughter’
cells. Bacteria can introduce variation into themselves by integrating additional DNA, often from their
surroundings, into their genome.
Bacteria can cause harm by directly invading and damaging tissue, others produce powerful chemicals known as
toxins, which damage cells. Antibiotics are commonly used to treat bacterial infections, but some strains of
Bacteria have become resistant to antibiotics, making them difficult to treat. Antibiotic resistance can be
transferred between bacterial strains through conjugation
Bacteria size will varies from 0.05–0.2 μm
 Bacteria

This is a common microscopic organism, which can multiply rapidly and build-up, causing
infection.
Most bacteria are harmless. Many types grow naturally on the human body and mucus
membranes.

Bacteria can be found on all workplace surfaces, in the soil and growing in various
substances used by workers.
Bacteria needs certain conditions to replicate, such as: Warmth, (5 to 60C), Moisture,
 Viruses
Viruses are different to bacteria as they are the smallest microbes.
Size of the viruses ranges from 20nm to 400 nm
A virus is a core genetic material, either Deoxyribonucleic acid (DNA) or Ribonucleic acid
(RNA), covered by a protective coat of protein.
Viruses replicate inside other living cells, they can latch on to host cells, taking over their
functions. The infected cell can infect new cells when stimulated.
Viruses are different to bacteria as they are the smallest microbes. A virus is a core genetic
material, either Deoxyribonucleic acid (DNA) or Ribonucleic acid (RNA), covered by a
protective coat of protein.
Viruses replicate inside other living cells, they can latch on to host cells, taking over their
functions. The infected cell can infect new cells when stimulated.
Viruses can cause several diseases in more advanced cell structures. Examples include:
chicken pox, cold, COVID 19, ebola, flu, hepatitis, herpes simplex virus (HSV), measles,
mumps, polio, rabies, rubella, smallpox.
Toxins from biological sources
Toxins are a subset of poison produced by living organisms. Poisons are any substance that can cause
harm to an organism if enough has been absorbed, this can be either through ingestion, inhalation, or
direct contact.
Many organisms produce toxins either as a defence mechanism or for predation, they tend to be
produced by bacteria, fungi, plants, insect, and animals.
The five most deadly toxins are:
botulinum toxin A, from the bacteria clostridium botulinum
tetanus toxin A, from the bacteria clostrifium tetani
diphtheria toxin, from the bacteria corynebacterium diphtheriae
muscarine, from the mushroom amanita muscaria
bufotoxin, from the common toad genus bufo.
Toxins can present in a variety of workplace settings. Venomous insects such as bees and wasps can
nest in any number of workplace buildings and can potentially sting workers. In many countries, snakes
and spiders are highly venomous, so can be a threat to workers.
 Anaphylaxis
A severe allergic reaction which can be fatal. This usually occurs in response to
almost any foreign substance. Common triggers include:
toxins from insect bites
Stings
Food
Medicines.
Anaphylaxis is a common medical emergency and a life-
threatening acute hypersensitivity reaction. It can be defined as
a rapidly evolving, generalized, multi-system allergic reaction.
Without treatment, anaphylaxis is often fatal due to its rapid
progression to respiratory collapse.
Long-term diseases, also called chronic diseases
Local diseases target one part (Lungs) of the body. An example of a local disease is hepatitis B (HBV). This can be
transmitted into the body through contact with infectious body fluids such as blood, vaginal secretions or semen.
Systemic diseases affect many parts of the body or the whole body. They can start as a local disease and progress to
systemic disease. For example, pneumonia may begin in one lung or both lungs but then spread throughout the body into
a potentially life-threatening condition.
Parasitic diseases are infectious diseases caused or transmitted by a parasite. A common example of parasitic disease is
toxoplasmosis. Infection usually occurs from:
eating undercooked contaminated meat, exposure to infected cat poo, or mother-to-child transmission in pregnancy.
Cancer
Carcinogens are substances that can cause cancer. Cancer is an uncontrolled growth of abnormal cells in the body. Some
new cases of cancer could be attributed to agents such as human papillomavirus (HPV), helicobacter pylori, hepatitis B and
C viruses.
Psychological conditions
There is a possible connection between infections and the development of disorders such as schizophrenia, depression and
bipolar disorder. A theory is that infection may influence the brain with infective agents, altering the central nervous
system.
Toxins (long-term)
Some toxins can have long-term effects. They affect people in different ways, from mild illness to death. For example, there
are several types of toxins produced by harmful algae, which in large quantities can form toxic blooms. These can be
responsible for causing:
respiratory irritation and distress, diarrhoea, vomiting, numbness, dizziness, paralysis, death.
Allergies
Allergies are long-term conditions that are not life threatening but cause localised tissue inflammation. Some biological
hazards can cause hypersensitivity, which is an over reaction by the immune system to an allergen. Examples include:
pollens from plants, viruses, bacteria, animals and birds.
 Biosafety levels
What Are Biosafety Levels?
Biological safety levels — are a series of protections specific to autoclave related
activities that take place in biological labs. Biosafety levels are individual safeguards
designed to protect laboratory personnel, as well as the surrounding environment
and community.
Each biosafety level — BSL-1 to BSL-4 — is defined based on the following:
Risks related to containment
Severity of infection
Transmissibility
Nature of the work conducted within the lab
Origin of the microbe
Agent in question
Route of exposure
 Biological safety cabinets (BSCs)
Biological safety cabinets (BSCs) are used to protect
personnel, products and the environment from
exposure to biohazards and cross contamination
during routine procedures.
BSCs are designed to handle hazardous pathogenic materials, among
other biohazards, and are used regularly in various types of
laboratories ranging from basic research to high containment.
Every BSC is categorized by a specific biosafety class: Class I, Class II
or Class III.
Biosafety Cabinet Class I
The Class I Biosafety Cabinet (BSC) provides personnel and
environmental protection, but no product protection.
It is similar in air movement to a chemical fume hood but has
a limited fixed work access opening, and the exhaust air must
be HEPA filtered to protect the environment.
However, to be classified as a Class I BSC, the inward flow of
air must be maintained at a minimum inflow velocity of 75
linear feet per minute (FPM) (0.38 m/s) through the front
access opening.
The Class I BSC is designed for general microbiological
research with low and moderate-risk agents or non-sterile
hazardous drug compounding in the pharmacy.
The Class I cabinet may be recirculated back into the
laboratory environment if no volatile chemicals a present.
A Class I CVE may be recirculated back into the pharmacy if
exhausted air passes through redundant (two) HEPA filters as
per USP 797 and USP 800.
 Class II cabinet

It is defined as a ventilated cabinet for personnel, product and environmental protection,

often used for microbiological work or sterile pharmacy compounding.
Class II BSCs are designed with an open front with inward airflow (personnel protection),
downward HEPA-filtered laminar airflow (product protection) and HEPA-filtered exhaust air
(environmental protection).
These cabinets are further differentiated by types based on construction, airflow and how
they interface with exhaust systems — A1, A2, B1, B2 and C1.
All Class II BSCs require all biologically contaminated ducts and plenums to be under negative
pressure or surrounded by negative pressure ducts and plenums. This provides a fail-to-safe
feature that protects the user even in the event of a plenum failure.
Type B2 cabinets take this a step further, requiring all biologically contaminated ducts and
plenums to be under negative pressure or surrounded by directly exhausted negative
pressure ducts and plenums.
Type C1 cabinets provide even more protection by maintaining containment from biological
and chemical hazards during building exhaust failures.
Class II, Type A2
A Class II, Type A2 biosafety cabinets are the most common type of BSC used today.
Must maintain a minimum average inflow velocity of 100 fpm through the sash opening.
Approximately 60% to 70% of the contaminated air is recycled and pushed back into the
workstation in the chamber through the downflow HEPA filter, while the remaining 30%
to 40% is exhausted through the exhaust HEPA filter.
The recirculated, HEPA-filtered downflow air (Laminar flow) creates an ISO 5
environment within the work area that protects the samples from external
contaminants.
When limited amounts of volatile chemicals are required, both NSF 49 and BMBL advise
the unit shall be connected to an external exhaust system via a canopy style exhaust
connection that will provide an audible and visual alarm within 15 seconds of an exhaust
failure.
It’s advised to never use a direct-connect or hard ducted connection when working with
hazardous chemicals. Canopy vented Type A2 cabinets may be used for work with
limited amounts of volatile chemicals if deemed appropriate by a chemical risk
assessment.
However, if hazardous, volatile chemicals are to be used within the cabinet, along with
the microbiological work, exhaust must be released into the atmosphere through the
direct duct system..
Class II, Type B1
Class II, Type B1 biosafety cabinets must maintain a minimum
average inflow velocity of 100 fpm through the sash opening
and must be connected to a building exhaust system.
They have HEPA-filtered downflow air composed mostly of
uncontaminated recirculated inflow air and exhaust most of the
contaminated downflow air through a dedicated duct after
passing through a HEPA filter.
Type B1 cabinets are safe for work involving limited amounts of
volatile chemicals and trace amounts of radionuclides, as long
as the work is done in the rear portion of the cabinet behind
the smoke split.
The rear portion is not marked or well-defined, and is ever-
changing as the BSC’s filters load, making this type of cabinet
unsafe for all but the most well-trained users.
Class II, Type B2

A Class II, Type B2 cabinets must maintain a minimum average inflow velocity
of 100 fpm through the sash opening and require a dedicated exhaust system
and dedicated remote blower for each cabinet.
All of the contaminated airflow (100%) in a Type B2 cabinet is externally
exhausted which means the air drawn into the cabinet is 100% exhausted into
the atmosphere.
They have HEPA-filtered downflow air drawn from the laboratory (not
recirculated from the cabinet exhaust) and exhaust all inflow and downflow air
out to the atmosphere after filtration through a HEPA filter.
Type B2 cabinets are suitable for work involving limited amounts of volatile
chemicals and trace amounts of radionuclides as an adjunct to microbiology
applications.
Although Type B2 BSCs offer protection when using higher volumes of chemicals
within the cabinet, they require complicated exhaust configurations and
consume large amounts of air to function.
As a result, Type B2 cabinets have the highest installation and operational costs
of any Class II BSC.
 Class III
A Class III cabinet is defined as a totally enclosed, ventilated cabinet with leak-
tight construction and attached rubber gloves for performing operations in
the cabinet.
These cabinets have a transfer chamber with interlocked doors that allow for
sterilization of materials before entering/exiting the glove box. Materials can
also be taken in and out through a dunk tank filled with a disinfecting
solution.
The cabinet is maintained under negative pressure and supply air is drawn in
through HEPA filters. The exhaust air is treated with either double HEPA
filtration or single HEPA filtration followed by air incineration and then
exhausted outside.
They are most commonly found in BSL 3 and BSL 4 laboratories, dubbed
cabinet laboratories.
Class IV

BSL-4. BSL-4 builds upon the containment

requirements of BSL-3 and is the highest level
of biological safety. There are a small number
of BSL-4 labs in the United States and around
the world. The microbes in a BSL-4 lab are
dangerous and exotic, posing a high risk of
aerosol-transmitted infections.
Biohazard Level 4 usually includes dangerous
viruses like Ebola, Marburg virus, Lassa fever,
Bolivian hemorrhagic fever, and many other
hemorrhagic viruses found in the tropics.
 Class IV
In addition to biosafety level 3 considerations, biosafety level 4
laboratories must follow these safety protocols:
Personnel must change clothing before entering the facility and shower
upon exiting
All materials must be decontaminated before leaving the facility
Personnel must wear the PPE from lower BSL levels, as well as a full-
body, air-supplied, positive pressure suit
Access to a Class III biological safety cabinet
BSL-4 labs are extremely isolated, often located in an isolated and
restricted zone of a building or in a separate building entirely.
BSL-4 labs also feature a dedicated supply of exhaust air, as well as
vacuum lines and decontamination systems.