BRM Training Manual

1. TRAINING MANUAL
ON
BIORISK MANAGEMENT
FOR LABORATORY WORKERS, FIELD PERSONELS AND RESEARCH
STUDENTS OF VETERINARY LABORATORIES
PAKISTAN BIORISK MANAGEMENT PROGRAM II
RELIEF INTERNATIONAL-PAKISTAN

2. PREFACE
Knowledge and awareness concerning Biorisk management is awfully inadequate in
Pakistan, for the most part, in regions of KPK and Balochistan provinces. whilst Relief
International analyzed numerous public veterinary hospitals and biological research
institutes, it was observed that biorisk is equally neglected in both veterinary and public
health sectors and workers, researchers and other related people are not only exposed to
this threat but worst, they do not realize the sensitivity of this issue. Relief International
conducted risk assessment of different hospitals and research institutes which served base
for development of this training manual and it is drafted for people working in such
facilities.
This manual has been developed to facilitate both the trainers and trainees to, not only, to
facilitate the conduction of training but also, will serve as, guideline notes for biorisk
management practices in laboratories, to be kept in facilities and confer with, every time,
obligatory after the training. It will serve the following categories of people
 Laboratory workers,
 Administration
 Researchers
 Support staff of hospitals and research institutes including doctors, assistants,
cleaning staff etc.
 Field workers and sample collection staff
The contents of this training manual will definitely increase the awareness level of related
persons and they will be able to deal pathological materials in a better way thus giving a
protection to not only to themselves but the environment as well. The administration can
benefit from this manual to draft principles and laws and practice at institutes to ensure
quality and protection of staff and better record maintenance, which will eventually
gratefully help to stop the spread of diseases.
Dr. Asghar Ali
National Program Manager
Relief International Pakistan.

3. ACRONYMS and ABBREVIATION
ABSA American Biological Safety Association
ABSL Animal Biosafety Level
AMP Assessment Mitigation Performance
BMBL Biosafety in Microbiological and Biomedical Laboratories
BRM Biorisk Management
BSC Biological Safety Cabinet
BSC Biosafety cabinet
BSL Biosafety Level
BSL Biosafety Level
BSL-3-Ag BSL-3-Agriculture
CCHF Crimean Congo Hemorrhagic Fever
CCTV Close Circuit Tele Vision
CDC Centers for Disease Control and Prevention
CDC Center of Disease Control
CFR Code of Federal Regulations
ELISA Enzyme-Linked Immunosorbent Assay
EPA Environmental Protection Agency
FDA Food and Drug Administration
FMD Foot and Mouth Disease
FMDV Foot and Mouth Disease Virus
GLP Good lap practices
GMO Genetically Modified Organism
HEPA High Efficiency Particulate Air
HVAC Heat ventilation and air conditioning
NIH National Institutes of Health
NIOSH National Institute for Occupational Safety and Health
OIE World Organization for Animal Health
OIE Office International des Epizootic
OSHA Occupational Safety and Health Administration
PCR Polymerase Change Reaction
PIN Personal Identification Number
PPE Personal Protective Equipment
SOP Standard Operating Procedure
TB Tuberculosis
USDA U.S. Department of Agriculture
UV Ultraviolet
VBM Valuable Biological Material
VS Veterinary Services

4. Biorisk Management
Contents:
 Background of BRM
 Scope and importance of BRM
 Objectives and importance of BRM training
 Terminology

Introduction
The risk associated with biological materials is called biorisk. Pathogens, biological
materials, diagnostic samples, culture material, strains, equipment, waste biological materials,
surfaces of tables in the lab that has the potential to communicate to laboratory workers,
researchers and environment. Similarly all Valuable Biological Materials (VBM) like Anthrax,
polio virus, H5N1 which has the fear to be stolen, misused or released intentionally from the lab
are the examples of biorisk.
Thousands of infectious biological agents and toxins are handled and processed in an
assortment of laboratory types for diagnostic, clinical, research, and commercial purposes around
the world. The type, number, and quantity of such materials are dependent upon the scope and
nature of the work conducted in the laboratory. Each agent and toxin handled is a potential
hazard posing a risk to personnel in the laboratory and facility, and likely to surrounding animal
and human communities beyond the laboratory.
Unfortunately at this time, in Pakistan, sufficient attention is not being paid to the safe
handling and disposal of biological specimens and cultures coming for analysis and diagnosis
that are capable of increasing the level of danger to the laboratory workers and simultaneously
spreading of infection, both to the community and environment. Thus it is the responsibility of
all laboratories/facilities that work with valuable biological material (VBM) to operate safely and
securely. The first step in achieving this operational goal is to assess the safety and security risks
present in these laboratories.
Biological agents and information developed to better health, welfare or safety could be
misused for harmful purposes to cause damage to public health, safety, and the environment.
Biorisk management describes containment principles, technologies, and practices to protect
people from biological agents, and prevent accidental release of biological agents. In addition to
biosafety, laboratory biosecurity measures aim to prevent theft and intentional or malicious use
of biological agents. Thus, both biosafety and biosecurity should be an integral part of program
management of facilities handling dangerous pathogens, in order to prevent, undesired spread,
theft or malicious use.

5. I. Scope and Importance of Biorisk Management:
a. The scope of biorisk management is to set requirements necessary to control risks
associated with the handling or storage and disposal of biological agents and toxins in
laboratories and facilities.
b. Establish and maintain a biorisk management system to control or minimize risk to
acceptable levels in relation to laboratory workers, the community and others as well
as the environment which could be directly or indirectly exposed to biological agents
or toxins.
c. Provide assurance that the requirements are in place and implemented effectively.
d. Seek and achieve certification or verification of the biorisk management system by an
independent third party.
e. Provide a framework that can be used as the basis for training and raising awareness
of laboratory biosafety and laboratory biosecurity guidelines and best practices within
the facilities.
II. Objectives of biorisk management training and training manual:
a. To make participants aware about laboratory biosafety and biosecurity protocols.
b. To sensitize and motivate them on the subject of laboratory biosafety and biosecurity
measures.
c. To enable them and encourage them to apply principles of biosafety and biosecurity
in facilities.

6. TERMINOLOGY
Biological laboratory:
A room within a facility, designated for work on biological agents and/or toxins
Biorisk:
The risk associated with biological materials. Examples are all pathogens, biological
materials, diagnostic samples, culture material, strains, equipment, and waste biological
materials. The work surfaces of tables and benches in the lab that have the potential to
communicate biorisk to laboratory workers, researchers and environment. Similarly all Valuable
Biological Materials (VBM) like Anthrax, polio virus, H5N1 which has the fear to be stolen,
misused or released intentionally from the lab.
Laboratory biorisk management:
System or process to control safety and security risk associated with handling or storage and
disposal of biological agents and toxins in laboratories and facilities.
Biorisk management is “the analysis of ways and development of strategies to minimize the
likelihood of the occurrence of biorisks”. 1
Laboratory biosafety:
It describes the containment principles, technologies, and practices that are implemented to
prevent the unintentional exposure to biological agents and toxins, and to prevent their accidental
release.
Laboratory biosecurity:
It describes the protection, control and accountability for biological agents and toxins within
laboratories, in order to prevent their loss, theft, misuse, diversion of, unauthorized access or
intentional unauthorized release.
Biorisk Assessment:
Process of identifying the hazards and evaluating the risks associated with biological agents
and toxins, taking into account the adequacy of any existing controls, and deciding whether or
not the risks are acceptable.
Biorisk mitigation:
Actions and control measures that are put into place to reduce or eliminate the risks
associated with biological agents and toxins.
Elimination and Substitution:
Removing the hazard, not working with the agent or replacing the hazard with something less
hazardous.
1
World Health Organization

7. Example: Replacement of liquid nitrogen storage in confined spaces by ultra-low temperature
storage cabinets (-135 ºC).
Water based chemicals rather than solvent based ones.
The use of a low pathogenic organism/strains rather than a highly pathogenic one.
Performance:
Implementation of the entire biorisk management system, including evaluating and ensuring
that the system is working the way it was designed.
Or another aspect of performance is the process of continually improving the system.
Control:
Continued procedures of monitoring for the laboratory work adopted by the laboratory staff
for releasing the reliable result.
Assurance:
Systematic process of checking through audits and inspections.
Improvement:
Setting and achieving management goals based on internal and external feedback.
Risk:
Likelihood of an undesirable event happening, that involves a specific hazard or threat and
has consequences.
Risk is a function of both the likelihood of something happening and consequences of that
occurrence. Or, more simply Risk = f (likelihood, consequences)
Examples: Risk with over speeding, pathogen, chemicals, gases and inflammable materials
Engineering controls:
Physical changes to work stations, equipment, materials, laboratory facilities or any other aspect
of the work environment that reduce or prevent exposure to hazards.
Examples: Biosafety cabinet, Positive pressure, CCTV (Close Circuit Tele Vision), badge reader
machine, Biometric, electronic barriers, animal containment caging system, autoclaves, sharps
containers and HEPA (High Efficiency Particulate Air) filter.
Administrative controls:
Policies, standards and guidelines used to control risks.

8. Practices and Procedures:
Processes and activities that is effective in reducing risks
1. To adopt SOPs during lab working
2. Good laboratory safety practices
Personal protective equipment:
Devices worn by the worker to protect against hazards in the laboratory such as gloves, shoe
covers, head cover, gown, safety glasses, mask or respirators etc.
BioRAM:
It is a computerized risk assessment tool developed by Sandia National Laboratory in
partnership with the international community to facilitate laboratory biosafety and biosecurity
risk assessments by simplifying risk characterization.
Incident:
An event with a potential for causing harm
Example: Needle pricks
Emergency:
An incident that requires an immediate response
Example: Electric shock, Fire hazard.
Contamination:
The presence of an infectious agent on a body surface; also on or in clothes, bedding, toys,
surgical instruments or dressings, or other inanimate articles or substances including water, milk,
and food, or that infectious agent itself.
Decontamination:
A process to remove contamination. Decontamination renders an area, device, item, or
material safe to handle, that is, reasonably free from a risk of disease transmission.
Sterilization:
A process that kills or removes all classes of microorganisms and spores or complete removal
of microorganism is called sterilization.
Disinfection:
A chemical or physical means of killing microorganisms but not necessarily spores

9. Antiseptic:
A substance that prevents or arrests the growth or action of microbes, either by inhibiting
their activity or by destroying them.
Example: Alcohol, iodine
Hazard:
Hazard may be a source, situation, or act with a potential for causing harm
1. Physical situation (e.g. a fire or explosion)
2. Activity (e.g. pipetting)
3. Biological material (biological agent or toxin and biological waste materials)
4. Chemicals and gases such as nitrogen
Biohazard:
Potential source of harm caused by biological agents or toxins
1. Viruses, bacteria, fungi, parasite
2. All blood, blood products, tissues and body fluids
3. Cultured cells (all human or animal) and potentially infectious agents.
4. Allergens
5. Toxins (bacterial, fungal, plant, etc.)
6. Clinical and diagnostic specimens
7. Infected animals, animal carcasses and animal tissues
Threat:
The possibility of an unfavorable event to occur, as an expression of target to cause evil,
injury, disruption or damage.
Adversary:
Individual with malicious intent or a person or group that is hostile to someone.
Example:
Conflict or dispute
Asset:
An item having value like Valuable Biological material in a laboratory or facility.
Valuable Biological Material (VBM):
Biological materials, that are the asset, required specific attention for protection and
monitoring in the lab and have the potential to cause harm in the population.

10. Example: VBM may include pathogens and toxins, vaccine strains, genetically modified
organisms (GMOs).
Standard operating procedure (SOP):
Set of written instructions that document a routine
or
Repetitive activity followed by an organization.
Characterization:
In characterization the property of the pathogens is determined such as whether the agent is
communicable or not, zoonotic potential of the agent, infectious dose, route of transmission
(inhalation, ingestion, skin contact), pathogenicity of agent, host range and epidemiology. Also
the likelihood of the risk and its consequences are assessed. Treatment and cost of treatment,
vaccine availability of the pathogen are also considered.
Example: Agent having property to communicate through inhalation has greater likelihood than
the one with ingestion or cutaneous mode of transmission. The Ebola virus has high likelihood,
high consequences as compared to anthrax or CCHF. Similarly TB has high likelihood and low
consequences as compared to CCHF.
Evaluation:
In the evaluation step the risk is estimated, quantified (quantitatively, qualitatively) using risk
matrix. This is a simple mechanism to increase visibility of risks and assist management decision
making. Identifying likelihood and occurrence and ordered them in low, moderate and high
levels.
Risk Matrix
It is a matrix that is used during risk assessment to define the various levels of risk as the
product of the harm probability categories (likelihood) and harm severity categories
(consequences).

11. Chapter 01:
Biorisk Assessment
Contents of chapter:
 Hazard and threat identification
 Hazard and threat analysis
 Level of biological risks
 Likelihood and consequences evaluation.

Hazard and Threat Identification
Hazard: It is synonym to danger, risk, chance of danger and chance of risk. It is defined as “Any
source of potential damage, harm or adverse health effects on something or someone under
certain conditions at work.
Or
A danger or source of danger with the potential to cause harm2
.
Examples: The sources of hazard could be a thing (knife), substance (chemicals), material
(pathogen), source (electric), condition (wet floor), process (culturing) and practices (pipetting).
lifesaving drugs at home as children can reach them, loaded gun that can fire, over speeding may
result in accident, room gas heaters may cause suffocation, use of mobile phone while driving
can resulted in accident.
Types of Hazard:
 Physical hazards (e.g. a fire or explosion)
 Chemical and gases hazards:
 Radiation hazards
 Biological hazards
2
Word Health Organization

12. What is biohazard?
Any Biological material which poses the potential to adversely affect the health of humans,
animals or environment is called biohazard.
Examples are:
1. Viruses, bacteria, fungi, parasite
2. Blood, blood products, tissues and body fluids
3. Cultured cells
4. Allergens
5. Toxins (bacterial, fungal, plant, etc.)
6. Clinical and diagnostic specimens
7. Infected animals and animal tissues
What is Threat?
The situation or substance when intends to create harm is referred as threat, e.g loaded gun in
the hands of stranger pointing to your head.
In biosecurity perspective threat is the intension of someone towards valuable biological
material to create harm to humans and environment. For example stealing of anthrax spores for
malicious intentions.
Hazard and Threat Analysis (frequency and magnitude of the hazard and threat)
The hazard or threat is the source or causative agent of a particular risk. The term hazard is
Used in the biosafety context and threat is used in the biosecurity context.
Hazard is the biological material worked with in the lab. For example if a biological agent is
highly communicable with inhalation route (hazard characteristic), and a procedure is being
implemented where large-volume aerosols are being generated in close proximity to the lab
worker, and protective equipment such as respirators or biosafety cabinets are unavailable (all
situation characteristics), we can say likelihood of exposure is high, and if that same biological
agent does not produce serious disease (hazard characteristic), and furthermore an effective
vaccine has been administered to all personnel (situation characteristic), we can say the
consequences of exposure are low.
The threat is the potential adversary who is interested in the biological materials.
Characterizing potential adversaries allows one to determine important parameters for likelihood
and consequences, such as means, motives, and opportunity.
Example: The use of physical barriers lowers the likelihood of threat. Availability of therapeutics
lowers the consequences of threat.
Training Manual on Biorisk Management Page 12

13. Hazard Analysis:
The hazard is evaluated by means of the following properties:
 Agent risk group classification (Risk group 1,2,3 and4)Routes of infection (
inhalation, ingestion and cutaneous)
 Infectious disease process ( incubation period)
 Virulence (the degree of pathogenicity).
 Pathogenicity ( ability of an organism to cause disease or to harm the host)
 Quantity, concentration, ( infectious dose, Lethal dose)
 Incidence in community ( outbreak)
 Presence of vectors ( Intermediate host like flies, ticks and mosquitos)
 Availability of preventative measures and effective treatment ( vaccines and antibiotics)
 Potential outcome of exposure ( morbidity and mortality)
 Stability of the agent in the environment ( thermo-stability)
Threat Analysis:
 Presence of valuable biological
material
 Presence of adversary in the area
(conflict, foreign invasion, insurgency
etc.)
 Level of training of staff
 Level of experience of staff
 Workload and fatigue
 Health and immune status of the
workforce
 Good laboratory practices
 Housekeeping and cleanliness
 Use of Biological Safety Cabinets
 Use of Personal Protective Equipment
 Proper Decontamination
 Waste Management

 Occupational Health
 Administrative Controls
 Presence of the Agent in the Environment
around the Laboratory
 Immune Status of the Community
 Population density of the Community
 Presence of suitable hosts or vectors


14. Levels of Biological Risks
What is risk?
Risk is the chance or probability that a person will be harmed or experience an adverse health
effect if exposed to a hazard.3
In other words we can also defined the risk as the possibility of undesirable event occurrence, if
happens can cause damage
Examples: risk with over speeding, pathogen, chemicals, gases and inflammable materials
What is Biorisk?
The probability or chance that a particular adverse event (accidental infection or unauthorized
access, loss, theft, misuse, diversion or intentional release) possibly leading to harm, will occur.4
OR
Biorisk is the likelihood of an undesirable event happening, that involves a specific bio hazard
or threat and has consequences.5
Risk = f (likelihood, consequences) or, more simply,
Risk is a function of both the Likelihood of something happening and consequences of that
occurrence
Examples are: all pathogens, biological materials, diagnostic samples, culture material, strains,
equipment, waste biological materials, surfaces of tables in the lab that has the potential to
communicate to lab: workers, researchers and environment. Similarly all Valuable Biological
Materials (VBM) like Anthrax, polio virus, H5N1 which has the fear to be stolen misused or
released intentionally from the lab.
The following risk group in relation with likelihood and consequences are: -
RGL1: No or low individual/community risk
A microorganism that is unlikely to cause human disease or animal disease.
Example: Escherichia coli K-12 strain.
RGL2: Moderate individual risk, low community risk
A pathogen that can cause human or animal disease but is unlikely to be a serious hazard to
laboratory workers, the community, livestock and environment is grouped in moderate level.
Laboratory exposures may cause infection, but effective treatment and preventative measures are
available and the risk of spread of infection is limited.
3
Word Health Organization
4
Word Health Organization
4 5
Sandia National Laboratories

15. Example: Salmonella species.
RGL3:High individual risk, low community risk
A pathogen that usually causes serious human or animal disease but does not ordinarily spread
from one infected individual to another. Effective treatment and preventive measures are
available.
Example: Mycobacterium tuberculosis
RGL4: High individual/community risk
A pathogen that usually causes serious human or animal disease and that can be readily
transmitted from one individual to another, directly or indirectly. Effective treatment and
preventive measures are not usually available.
Example: Ebola virus
Likelihood and Consequences Evaluation
Likelihood is the probability of something to happen. To evaluate the likelihood we have to
consider the agent, the procedures happening at the laboratory and the mitigation measures in
place. The same could be explained as below:
Risk Group Individual Risk Community Risk
RG-One Low No
RG-Two Medium Low
RG-Three High Medium
RG-Four High High
Properties of Agent Laboratory Procedures Biosafety Measures
 Infectious  Culture/isolation  Engineering controls

routes  Pipetting  Bio Safety Cabinet (BSC)
Infectious dose  Injecting  Sealed flasks
 Stability  Diagnosis  HEPA filtered exhaust
 ELISA  PPE
 PCR  Gloves, respirators, etc.
 Administrative Controls
 Policies and Procedures
Any increase and decrease in the above all will alter the likelihood, for example if the hazard
has inhalation route of transmission the likelihood will increase, but the same hazard if
manipulated in the biosafety cabinet the likelihood will decrease and vice versa.

16. Consequences refer to the impact of the pathogen on human and environment. This can be
evaluated by the mean of understanding the agent, vulnerability of the community, availability of
therapeutics and prophylaxis etc.: -
Agent: Properties of the agent, route of transmission of the agent, virulence, pathogenicity,
stability outside the host, transmission between humans and animals, mortality, morbidity, case
fatality rate. In addition of the above, general health status of the community, prophylaxis
availability, treatment availability for the specific agent.
For further study and reference:
http://www.who.int/csr/resources/publications/biosafety/WHO_CDS_EPR_2006_6.pdf
http://www.biosecurity.sandia.gov/ibtr/subpages/pastConf/20102011/absa/handouts.pdf

17. Chapter 2:
Biorisk Mitigation
Contents:
 Introduction to biorisk mitigation
 Elimination and substitution
 Engineering Control
 Administrative Control
 Practices and Procedures
 Personal Protective Equipment
Introduction to Bio-risk Mitigation
BRM has three main components Assessment, Mitigation and Performance
BRM= (Assessment) + (Mitigation) + (Performance) or BRM=AMP
Scientifically BRM can be defined as System or process to control biosafety and biosecurity
risks associated with the handling or storage and disposal of biological agents and toxins in
laboratories and facilities.
Three important definitions:
Risk: The chance (probability or possibility) of exposure to danger/damage/injury/loss
Hazard: Is a source that has a potential to cause harm.
Threat: A person or thing likely to cause damage or danger.
Mitigation: The action of reducing the severity, seriousness, or painfulness of something6
Definition of bio-risk mitigation:
Actions and control measures that are put together to eliminate or reduce the risks associated
with biological agents and toxins.7
Or
A process through which we reduce risk that were identified during the risk assessment to
ensure biosafety and biosecurity
6
Oxford Advanced Learner's Dictionary
7
Sandia National Laboratories

18. Mitigation is based on the risk evaluated and the degree of risk. The prime objective of the
mitigation is to ensure biosafety of individual, population, environment and biosecurity of
valuable biological material (VBM).
Types:
All the mitigation measures come under the following types or classes of biorisk mitigation.
- Elimination and Substitution
- Engineering Controls
- Administrative Controls
- Practices and Procedures
- Personnel Protective Equipment
Elimination and Substitution
Elimination: Elimination is the decision to remove the risk:
 Killing of rabid dogs.
 Culling of bird flu affected flock or eradication of cows in case of mad cow disease.
 Not working with CCHF in the laboratory with BSL2 facilities.
Substitution: The action of replacing someone or something with another person or thing or in
biorisk management it is the replacement of more hazardous with less hazardous.
 Replacing a window Air conditioner by split one in the room.
 Manual gear in car replaced by automatic one.
 E-tag instead of manual collection in a tool plaza.
 Use of low pathogenic organisms rather than high pathogenic organisms.
 Using Nitrogen cylinders for semen storage instead of ultra-low freezer.
 Replacing mouth pipetting with automatic.
Elimination or substitution is the most effective means of mitigation
Engineering Control
Engineering controls are the physical changes that are brought to work station, equipment,
materials and production facilities to reduce or prevent exposure to biohazard.
Example:
 Biological safety cabinets, autoclaves, sharps containers, PPE
 Facility ventilation and air flow

 Physical security: access restrictions, CCTV camera, facility and equipment locks with
associated key control protocols, badge readers, detectors and sensors and biometric
devices.



19. Administrative Control
Administrative controls are the policies, standards and guidelines that are used to mitigate risk. It
includes,
 Qualified, trained, and competent personnel
 Hierarchy well defined terms of reference, accountability, check and balance,
implementation of standard operating procedures.
 Health and safety programs e.g. Vaccination
 Emergency response and contingency plans

- Precautions against natural disasters
- Provision of emergency equipment (protective clothing, disinfections)
- Identification of responsible persons and their duties
- Shifting of expose or infected persons to hospital in case of emergency.
- Emergency treatment of exposed or injured persons
 Biological agents and toxin inventory management requirements

- Record of date received on each chemical container.
- Maintenance of inventory of chemicals stored in lab.
- Annual update of inventory list.

 Waste management policies

- Waste storage (containers, colour coded bins, plastic bags )
- Waste management handling, transportation and disposal
 Security policies
- Facility security
- Visitor access
- Personnel security
- Access to biological agents and toxin
- Data security
Practices and Procedures
Processes and activities that are practiced to reduce biorisk
Examples:
 Standard operating procedures for all safety and laboratory biosecurity relevant processes
 Good laboratory safety practices

- Training before starting work in the laboratory
- No eating and drinking

20. - Use safety cabinets
- Proper labeling of lab equipment
- Location of emergency exits
- SOPs for each test /equipment must be displayed
- Name tags for every person working in the lab
- Unauthorized person must not to enter the lab
 Use of appropriate disinfection and decontamination practices

- All items in the Biosafety Cabinets including equipment should be decontaminated
before and after use
- The work surface and interior walls must be wiped that must kill microorganism.
 Transportation procedures for infectious materials and specimens

- Packing
- Marking
- Labeling
 Specimen and reagent handling and storage practices

- Ensure that everything required to take sample is accessible
- Specimen and reagents should be properly labeled
- The head of the container must be properly screwed to avoid any leakage
- Persons handling the specimens or reagents must use proper PPE.

 Emergency exercise drills.

- Practices drills should address a variety of scenarios such as loss or theft of materials
- Emergency response to accidents, injuries, incidents reporting and identification of
and response to security breaches.
- Firing
 Training on the safe and secure handling of biological agents and toxins

- Never mouth pipette
- Avoid hand to mouth or hand to eye contact in the lab.
- Never eat or drink in the lab
- Never use cosmetics in the lab including chap stick and wearing of lens
- Use aseptic techniques
- Hand washing after removal of gloves and other personnel protective equipment
- Clean laboratory work surface with disinfectants after working with infectious
materials.

22. Personnel Protective Equipment (PPE)
Devices or gear worn by the worker to protect against hazards in the laboratory are known as
personal protective equipment.
 Eye protection (safety glasses, face shields)
 Respiratory protection (face masks, N95 respirators, face shields)
 Hand protection (gloves)

 Body protection (laboratory coats, uniforms, coveralls, surgical scrubs, rubber boots,
disposable shoe covers)
For further study and references:
1. http://www.biosecurity.sandia.gov/ibtr/subpages/pastConf/20102011/abot/mitigation.pdf
2. http://www.who.int/ihr/training/6a_Session3.pdf
3. Handbook of Laboratory Biorisk Management by Reynolds M. Salerno Publisher Taylor and Francis,
2013.
4. Biological Laboratory Applied Biosecurity and Biorisk Management Guide: Developing, Creating
and Sustaining an Applied Biorisk Program by Robert Hawley, J. Craig Reed Publisher Kirk R
Wilhelm, 2015.

23. Chapter 03:
Performance
Contents:
 Introduction to Performance
 Key Elements of Performance
 Control ( monitoring protocols)
 Assurance
 Improvement

Introduction to Performance
Performance: Implementation of the entire Biorisk management system including evaluating
and ensuring that the system is working the way it was designed. Another aspect of performance
is the process of continually improving the system
 Is the way in which someone or something functions
 It improves biorisk Management
To know that system works and is sustainable and that the risk is acceptable.
How Performance Improve BRM?
Performance has the following parts.
1. Control- Whether system works proper
2. Assure - Whether system is sustainable
3. Improve - Whether the risk is acceptable
1. Control: Control is continues procedures of monitoring for the laboratory work adopted by
the laboratory staff for releasing the reliable result. To manage biorisk, the following steps
should be considered
Steps in Control
1. Policy (what to do)
E.g. policies for sample record, visitor record, use of PPE, proper waste management,
collection point (sample and report), documentation.
2. Procedure (how to do it) (Standard Operating Procedures)
E.g. Manuals, SOPs, design records, inspection checklists, training records, etc.
3. Structure (Organization structure)
E.g. hierarchy, Director, principal in-charge, Laboratory technicians,
4. Responsibility (who to do, what and when) roles, responsibilities and authorities.
E.g. Lab technicians/trained staff for sample diagnostic, administrator or monitor for
SOPs and policy, biosafety committee for biosafety protocol.

24. 2. Assure: Systematic process of checking through audits and inspections.
This process can be done on weekly, monthly or quarterly basis to check the results and
impacts of the actions taken for Bio-risk Management through pre and post-analytic phase, test
standardization, performance evaluations (internal evaluation or external evaluation), accident /
incident investigation, inspection and audit. Senior leadership reviews of the Bio-risk
management system at planned intervals to ensure its continuing suitability, adequacy and
effectiveness. The review shall include assessing opportunities for improvement and the need for
changes to the system, procedures, policies and objectives. Records from the management
review shall be maintained. The management review should be conducted at a defined frequency
determined by the needs of the organization, but at least annually.
The review input should include information on:
 Results of audits;
 Compliance to standard operating procedures and work instructions;
 Status of risk assessment activities;
 Status of preventive and corrective actions;
 Follow-up actions from previous management reviews;
 Changes that could affect the system;
 Recommendations for improvement;
 Results of accident / incident investigations.
The review output should include decisions and actions related to improvement.
3. Improvement
Setting and achieving management
i) Improvement of the effectiveness of the Bio-risk Management System
ii) Improvement related to the requirements and risk assessments
iii) Resource needs.
iv) Goals based on internal and external feedback
Systematic Approach
Assessment = Plan, Do, Check, Act
Mitigation = Plan, Do, Check, Act
Performance = Plan, Do, Check, Act

25. Mitigation is improved and sustained when performance measures are included
1. Pre and post-analytic phase: Three phases of laboratory testing: Pre-analytical
specimen collection, transport and processing
2. Analytical—testing
3. Post-analytical—testing results transmission, interpretation, follow-up, retesting.
Test standardization: Test Standardization is achieved when test results with the same high
levels of accuracy and care can be reproduced across measurement systems, laboratories, and
over time. Standardization depends on a careful process using:
 Reference materials
 Proficiency testing
 Training
 Guidelines
 Consultations
Accident / Incident Investigation: The laboratory shall establish and maintain documented
procedures to define, record, analyze and learn from accidents and incidents involving biological
agents and toxins.

26. The purpose of an incident investigation is to find out what happened during the incident,
why it happened, and determine what steps should be taken to prevent a recurrence. To be
effective, the active participation of supervisors and workers is essential.
Inspection and Audit: The organization shall ensure that a program of inspection and audit is
conducted which is appropriate to the risk associated with the facility.
Inspections and audits shall be conducted at planned intervals to determine if the bio-risk
management system confirms to the documented plans and to the requirements of this
agreement, and that it is effectively implemented and maintained.
Management responsible for the area being inspected / audited shall ensure that any action is
taken without undue delay to eliminate detected non-conformities and their causes.

27. Chapter 04:
Biosafety Introduction
(The facilitator can apply brainstorming, class room discussions, role plays, demonstration,
group activities and question and answer techniques to introduce the course participants with
biosafety and different biosafety levels. The facilitator can use different teaching aids like white
board, sticky notes, charts, diagrams, demonstration materials and power point presentations in
the session depending upon the session plan. The facilitator will develop session plan for each
session keeping in view level of participants, level of awareness regarding biorisk management,
existing skills and attitude towards biorisk management).
Contents:
 Introduction to biosafety
 Biosafety level I
 Biosafety level II
 Biosafety level III
 Biosafety level IV

Introduction to biosafety
A set of preventive measures designed to reduce the risk of accidental exposure to biological
hazard. The application of knowledge, techniques and equipment to prevent personal, laboratory
and environmental exposure to potentially infectious agents or biohazards is called biosafety.
Biosafety defines the containment conditions under which infectious agents can be safely
manipulated. The objective of containment is to confine biohazards and to reduce the potential
exposure of the laboratory worker, persons outside of the laboratory, and the environment to
potentially infectious agents.
Purpose of biosafety
The purpose of biosafety is to establish a prevention barrier to disease causing agents and
other threats i.e. protective clothing, protective equipment, biosafety cabinets, automatic pipette
etc
Biosafety requirement
 Conducting risk assessments to identify and control workplace safety and health risks.
 Providing safe work facilities and arrangements for their workers;
 Ensuring safety in machinery, equipment, substances and work processes at the
workplace;
 Providing adequate instruction, information, training and supervision to workers
 Implementing control measures for dealing with emergencies
 Guidelines developed to protect workers in microbiological and medical labs
through engineering controls, management policies, and work practices.

28. Biosafety levels
Bio Safety Levels (BSL) refers to a quarantined room/facility which a virus or bacteria is
contained in.
So, based on the degree of hazard posed by these agents, labs are divided into four biosafety
levels, and mandated protective practices increase with each level
Objectives of classification
 Define barriers and procedures used by laboratories to protect workers and others from
infection.
 Describe the protective measures used by each type of laboratory when handling
infectious materials.
 Describe types of biological agents handled in each type of laboratory.
 Identify typical places where each type of laboratory can be found.
 Classify the labs accordingly.

29. Levels of classification
Biosafety levels for Labs define that work with infectious agents and determine:
 Regulations
 Outline
 Precautions
 Special practices
 Decontamination procedures
Note: In any laboratory, precautions must be taken so that the people researching or trying to
identify organisms do not become infected themselves. According to the Centers for Disease
Control and Prevention (CDC), scientists and lab technicians have to be very aware of
Microorganisms; while handling or testing clinical specimens, they could accidentally infect
themselves or their coworkers. Because of the danger, labs must adhere to very specific safety
regulations to work with organisms that pose a hazard to human health. Protective practices
increase with each level of biosafety.
The regulations outline precautions, special practices, and decontamination procedures for
labs that work with infectious agents. Based on the degree of hazard posed by these agents, labs
are divided into four biosafety levels and mandated protective practices increase with each level.
Biosafety Level 1 labs work with the least dangerous agents and require the fewest precautions;
Biosafety Level 4 labs have the strictest methods for handling organisms because they deal with
agents that are most dangerous to human health.
Each biosafety level has prescribed barriers to protect against microorganisms. Primary
barriers are physical barriers or personal protective equipment between the lab worker and the
pathogen, such as gloves, masks, or special breathing apparatuses.
Secondary barriers are structural aspects of the laboratory itself that make the working
environment safer against the risk of infection; these include sinks for hand washing, special
containment areas for working directly with organisms and special air ventilation patterns
designed to prevent contamination of other rooms and other workers in the building.
Universal Precautions:
Biosafety precautions are important outside the lab also. Universal precautions have been
developed to protect health professionals. While these precautions most often apply in a clinical
setting such as a hospital or doctor’s office, they may also be important for field epidemiology
practices during an outbreak investigation, particularly when collecting lab specimens.
Universal precautions include hand hygiene, use of gloves, gown, masks, eye protection, and
face shields, and safe injection practices. Universal precautions also require that all equipment or
items likely to have been contaminated with infectious fluids are handled in a manner that
prevents transmission of any infectious agents.
Special circumstances such as the decontamination of methamphetamine labs may require
additional precautions such as protective clothing and special site decontamination.

30. 5.3 Biosafety level 1 (BSL – 1)
 Space and facilities should be provided for the safe handling and storage of solvents,
radioactive materials, and compressed and liquefied gases.

 Facilities for storing outer garments and personal items should be provided outside the
laboratory working areas.

 Facilities for eating and drinking and for rest should be provided outside the laboratory
working areas.

 Hand-washing basins, with running water if possible, should be provided in each
laboratory room, preferably near the exit door.

 Doors should have vision panels,and preferably be self-closing.
Agents:
Agent not known to always cause diseases in healthy adults or a microorganism that is
unlikely to cause human or animal disease.
E.g. Bacillus subtilis, Non pathogenic Eschericia coli species
Practices:
Standard practices required;
 Frequent hand washing
 Door that can be kept closed when working
 Limits on access to the lab space when working
 No smoking, eating, drinking, storage of food in laboratory
 Care to minimize splashes and actions that may create aerosols (tiny droplets)
 Decontamination of work surfaces after every use and after any spill
 Decontamination of laboratory wastes
 Use of mechanical pipettes only (no mouth pipetting)
 "Sharps" precautions, including special containers for disposing of needles and other
sharp objects;
 Maintenance of insect/rodent control program
 Use of personal protective equipment (lab coats, latex gloves, eye protection or face
shields)
 Open bench top sink for hand washing. No other major facility structures are required for
BSL-1 labs
Example: High schools/collages lab, Drinking water treatment facility

31. 5.4 Biosafety level 2 (BSL – 2)
Procedures likely to generate aerosols are performed within a biological safety cabinet.
Doors are kept closed and are posted with appropriate hazard signs. Potentially contaminated
wastes are separated from the general waste stream.
 At Biosafety Level 2, an autoclave or other means of decontamination should be
available in appropriate proximity to the laboratory.
 Safety systems should cover fire, electrical emergencies, emergency shower and eyewash
facilities.
 First-aid areas or rooms suitably equipped and readily accessible should be available
 In the planning of new facilities, consideration should be given to the provision of
mechanical ventilation systems that provide an inward flow of air without recirculation.
If there is no mechanical ventilation, windows should be able to be opened and should be
fitted with arthropod-proof screens.
 A dependable supply of good quality water is essential. There should be no cross
connections between sources of laboratory and drinking-water supplies. An anti backflow
device should be fitted to protect the public water system.
 There should be a reliable and adequate electricity supply and emergency lighting to
permit safe exit. A stand-by generator is desirable for the support of essential equipment,
such as incubators, biological safety cabinets, freezers, etc., and for the ventilation of
animal cages.
 Laboratories and animal houses are occasionally the targets of miscreants. Physical and
fire security must be considered. Strong doors, screened windows and restricted issuance
of keys are compulsory. Other measures should be considered and applied, as appropriate
to increase security.
Agents:
Agents associated with human diseases routes of transmission include percutaneous injury,
ingestion, and mucous membrane exposure.
E.g. Vaccine strains of virus, human/animal blood and blood products, Herpes simplex
viruses, HIV, Clostridium botulinum, Clostridium tetani, Campylobacter fetus, coli,
Corynebacterium, Entamoeba, Cryptosporidium, Giardia spp, Plasmodium spp.
Primary hazards:
In working with BSL-2 agents, the primary hazards to personnel are accidental needle pricks,
potential infection through exposure to the eyes and nose (mucous membranes), and ingestion of
infectious materials.
BSL-2 agents do not cause lethal infections and are not transmissible via the airborne route.
This means that they do not cause infection if tiny droplets of the material become airborne (i.e.,
aerosolized) and are inhaled, which might occur if the material were spattered Accidental needle
pricks
 Exposure to eyes and nose (mucous membranes)
 Ingestion of infectious materials


32.  Agents do not cause lethal infections, are not transmissible via airborne route
 (Do not cause infection if tiny droplets become airborne and are inhaled, which might
occur if the material were spattered)
 Agents are pathogens for which immunization or antibiotic treatment is available
 Extreme care should be taken with contaminated needles and sharp lab instruments
Standard practices include BSL-1 plus:
To reduce accidental infection, special procedures for BSL-2 labs include all standard
practices for BSL-1 labs, plus a few others as following:
 Special policies and procedures to restrict access to the lab when work is being
conducted.
 Biohazard warning signs posted outside the lab
 Surveillance of laboratory personnel with appropriate immunizations offered.
 A biosafety manual that includes definitions of any needed waste decontamination or
medical surveillance policies specific to the activities and agents in that lab.
 Supervisory staff that have experience in working with infectious agents and specific
training for laboratory personnel in handling these agents.
Primary barriers: biosafety cabinets or other approved containment devices
Some primary barriers in BSL-2 labs are biosafety cabinets or other approved containment
devices. These areas minimize potential contamination while working with an agent, particularly
if there may be splashes or aerosolization of infectious materials.
Personal protective equipment includes lab coats, gloves, and face protection as needed when
working with infectious agents. Protective clothing must be removed when personnel leave the
laboratory area.
Cabinets should be thoroughly decontaminated daily and if radioactive materials are used,
monitored for radiation as a method of personal protection.
Secondary barriers include all BSL-1 barriers, plus an autoclave (sterilization machine) for
lab glassware.
 Personal protective equipment: lab coats, gloves, face protection as needed
 Protective clothing removed when personnel leave laboratory area
 Cabinets thoroughly decontaminated daily and monitored for radiation for personal
protection.
Secondary barriers: BSL-1 barriers plus autoclave for glassware
Facilities (secondary barriers):
Secondary barriers include all BSL-1 barriers, plus an autoclave (sterilization machine) for
lab glassware.
Examples: local health/veterinary departments, universities, private clinical diagnostic
laboratories.

33. Biosafety level 3 (BSL – 3)
The laboratory is separated from general traffic flow and accessed through an anteroom
(double door entry or basic laboratory of Biosafety Level 2) or an airlock. An autoclave is
available within the facility for decontamination of wastes prior to disposal. A sink with hands-
free operation is available. Inward directional airflow is established and all work with infectious
materials is conducted within a biological safety cabinet.
A BSL-3 laboratory must be used when work is done with indigenous or exotic agents that
have the potential for respiratory (aerosol) transmission and may cause serious and potentially
lethal infections. Materials potentially infected with these agents may be studied at a BSL-2 level
for diagnostic purposes only, but further manipulation and experimentation require BSL-3
conditions.
Agents studied in a BSL-3 lab include Mycobacterium tuberculosis Encephalitis virus, and
Coxiella burnetii.
Primary hazards: needle pricks, ingestion, exposure to infectious aerosols.
The containment laboratory - Biosafety Level 3 is designed and provided for work with Risk
Group 3 microorganisms and with large volumes or high concentrations of Risk Group 2
microorganisms that pose an increased risk of aerosol spread. Biosafety Level 3 containment
requires the strengthening of the operational and safety program over and above those for basic
laboratories - Biosafety Levels 1 and 2.
The guidelines given in the form of additions to those for basic laboratories - Biosafety
Levels 1 and 2, which must therefore be applied before those specific for the containment
laboratory - Biosafety Level 3. The major additions and changes are in:
 Code of practice
 Laboratory design and facilities
 Health and medical surveillance.

Laboratory Equipment:
The principles are same as for the basic laboratory - Biosafety Level 2. However, at
Biosafety Level 3, manipulation of all potentially infectious material must be conducted within a
biological safety cabinet or other primary containment device like centrifuges

34. Practices:
BSL-2 practice plus:
 Strictly controlled access to the lab.
 Decontamination of all waste and decontamination of laboratory clothing before
laundering.
 Specific training for lab personnel in handling potentially lethal agents.
 Changing contaminated protective lab clothing, decontaminating lab clothing before
laundering.
 Institutional policies regarding specimen collection and storage from workers to establish
exposure.
Primary barriers and safety equipment:
Primary and secondary protective barriers in the BSL-3 lab emphasize protecting lab
personnel, as well as personnel in nearby lab areas, the community, and the environment from
exposure to potentially infectious aerosols. Primary barriers are similar to BSL-2 personal
protective equipment, but may also include respiratory equipment if there is a risk of infection
through inhalation.
Secondary barriers at BSL-3 labs include all BSL-2 barriers, plus a few more sophisticated
barriers.
 Corridors must be separated from direct access to the laboratory.
 Access must be through self-closing double doors.
 Air handling systems must be designed to ensure negative air flow, so that air around
doors and windows flows into the laboratory rather than out of the laboratory.
 Air pumped into the laboratory is not re-circulated in the building. This measure is to
prevent infectious aerosols from being carried outside the lab through the air.
 PPE: Protective laboratory clothing, gloves, face, eye and respiratory protection, as
needed.
Laboratory Design and Facilities:
The laboratory design and facilities for basic laboratories - Biosafety Levels 1 and 2 apply except
where modified as follows:
 The laboratory must be separated from the areas that are open to unrestricted traffic flow
within the building. Additional separation may be achieved by placing the laboratory at
the blind end of a corridor or constructing a partition and door or access through an
anteroom (e.g. a double-door entry or basic laboratory-Biosafety Level 2), describing a
specific area designed to maintain the pressure differential between the laboratory and its
adjacent space. The anteroom should have facilities for separating clean and dirty
clothing and a shower may also be necessary.

 Anteroom doors may be self-closing and interlocking so that only one door is open at a
time. A break-through panel may be provided for emergency exit use.


35.  Surfaces of walls, floors and ceilings should be water-resistant and easy to clean.
Openings through these surfaces (e.g. for service pipes) should be sealed to facilitate
decontamination of the room(s).

 The laboratory room must be sealable for decontamination. Air-ducting systems must be
constructed to permit gaseous decontamination.
 Windows must be closed, sealed and break-resistant.

 A hand-washing station with hands-free controls should be provided near each exit door.

 There must be a controlled ventilation system that maintains a directional airflow into the
laboratory room. A visual monitoring device with or without alarm(s) should be installed
so that staff can, ensure that proper directional airflow is maintained into the laboratory
room at all times.

 The building ventilation system must be constructed that air from the containment
laboratory – (Biosafety Level 3) is not re-circulated to other areas within the building. Air
may be high-efficiency particulate air (HEPA) filtered, reconditioned and re-circulated
within that laboratory. When exhaust air from the laboratory (other than from biological
safety cabinets) is discharged to the outside of the building, it must be dispersed away
from occupied buildings and air intakes. Depending on the agents in use, this air may be
discharged through HEPA filters. A heating, ventilation and air-conditioning (HVAC)
control system may be installed to prevent sustained positive pressure of the laboratory.
Consideration should be given to the installation of audible or clearly visible alarms to
notify personnel of HVAC system failure.

 All HEPA filters must be installed in a manner that permits gaseous decontamination and
testing.

 Biological safety cabinets should be sited away from walking areas and out of cross-
currents from doors and ventilation systems.

 The exhaust air from Class I or Class II biological safety cabinets, , must be discharged in
such a way as to avoid interference with the air balance of the cabinet or the building
exhaust system.

 An autoclave for the decontamination of waste material should be available in the
containment laboratory. If infectious waste has to be removed from the containment
laboratory for decontamination and disposal, it must be transported in sealed,
unbreakable and leak proof containers according to national or international regulations,
as appropriate.

 Backflow-precaution devices must be fitted to the water supply. Vacuum lines should be
protected with liquid disinfectant traps and HEPA filters or their equivalent. Alternative
vacuum pumps should be properly protected with traps and filters.

 All facility design and operational procedures should be documented.

36. Facilities (secondary barriers):
BSL-2 plus:
 Physical separation from access corridors, Self-closing, double-door access.
 Exhausted air not re-circulated.
 Negative airflow into laboratory.
 Entry through airlock or anteroom.
 Hand washing sink near laboratory exit.
[
Biosafety level 4 (BSL – 4)
The maximum containment laboratory - Biosafety Level 4 is designed for work with Risk
Group 4 microorganisms. Before such a laboratory is constructed and put into operation,
intensive consultations should be held with institutions who had experience of operating a
similar facility. - Biosafety Level 4 should be under the control of national or other appropriate
health authorities.. Entities working to pursue development of a Biosafety Level 4 laboratory
should contact the WHO Biosafety program for additional information.
Agents:
 Dangerous/exotic agents which post high individual risk of aerosol-transmitted laboratory
infections that are frequently fatal, for which there are no vaccines or treatments.

 Agents with a close or identical antigenic relationship to an agent requiring BSL until
data are available to re-designate the level.

 Related agents with unknown risk of transmission.
 All BSL 4 agents are viruses.
E.g. Congo-Crimean hemorrhagic fever, Ebola

38. About the Information:
1. http://www.cdc.gov/od/ohs/biosfty/bmbl4/ bmbl4s6.htm
2. World Health Organization (WHO) Biosafety Manual Third Edition.

39. Chapter 05:
Good Laboratory Practices
Contents:
 Importance of Good Lab Practices ( GLPs)
 Good Lab work practices
 Who is responsible for enforcing GLPs?
 Barriers to good laboratory practices
 Laboratory equipment validation
Importance of Good Lab Practices (GLPs)
Good laboratory practices (GLP) are techniques and methods of doing work in the laboratory
that reduce biorisk. Good laboratory practices (GLP)) not only reduce risk but also promote
better research, more accurate results and better data. Good laboratory practices (GLP) can be
enforced/ promoted through both administrative (e.g. policy, SOPs) and engineering controls
(e.g. personal protective equipment, biosafety cabinet, door locks, fridge lock, etc)
"Good Laboratory Practice (GLP) is concerned with the organizational process and the
conditions under which laboratory studies are planned, performed, monitored, recorded, and
reported."8
“Good lab practices are working methods applied to eliminate or minimize exposure to
biological agents via e.g. aerosols, splashes and accidental inoculation”9
A Good Laboratory Practice (GLP) is a practice, technique, or procedure that, when followed
or demonstrated, will result in protection of lab workers,the environment and minimizing the risk
of exposure to hazardous agents. GLP prescribes a laboratory to work according to a system of
procedures and protocols. GLP is a policy for all aspects of the laboratory which influence the
quality of the analytical work. When properly applied, GLP should then:
 Allow better laboratory management (including quality management).
 Improve efficiency (thus reducing costs).
 Minimize errors.
 Allow quality control (including tracking of errors and their cause).
 Stimulate and motivate all personnel (well defined roles and responsibilities of staff,
provision of all required personal protective equipment to staff , make easy their work
and reduce stress)
 Improve safety
 Improve internal and external channels of communication.
An important aspect is also that the standards of quality are documented and can be
demonstrated to authorities and clients. This results in an improved reputation for the laboratory
and for the institute as a whole.
8
OECD (Organization for Economic Cooperation and Development)
9
CWA 15793 ( CEN Workshop Agreement)

40. 6.2 Good laboratory practices
The following are major good lab practices
1. Orientation of lab workers, researcher students about the work.
2. No eating drinking and mouth pippetting in the laboratory.
3. Use of appropriate safety cabinet.
4. Cleanliness: Cleaning procedures shall include at least
• Responsibility for cleaning
• Item/area to be cleaned
• Frequency of cleaning
• Method of cleaning, including dismantling equipment for cleaning purposes where
required
• Cleaning chemicals and concentrations
• Cleaning materials to be used
• Cleaning records and responsibility for verification
5. Spill cleaning as per standard requirement
6. Hand washing before and after work
7. Proper disposal of laboratory waste materials
8. Use of sterile material/equipment for each patient/sample/work
9. Proper labeling of laboratory equipment/chemicals/wastes
10. Location of emergency exits in laboratory
11. Displaying of signs and symbols in the facility
12. Maintenance of laboratory record/ ledger book /inventory/documentation
13. Standard operating procedures for each test/equipment should be displayed
14. Use of appropriate personal protective equipment
15. Unauthorized persons not allowed to the laboratory
16. Pregnant women / children not allowed in the laboratory
17. Name tags for laboratory workers.
6.3 Who is responsible for enforcing GLP?
National/International Laws (The United States FDA (Food and drug administration) has
rules for GLP in 21CFR58.) (Veterinary Medicinal Products; Directive 2001/82/EC of the
European Parliament and of the Council of 6 November 2001 on the Community code relating to
veterinary medicinal products) (OECD: EMV/MC/CHEM (98)17 part two).
 Institution: Institutional Administration and Biosafety Committee
 Laboratory: Laboratory manager/supervisor, laboratory staff
 Everyone: It is everyone’s responsibility to uphold GLP


41. 6.4 Barriers to good laboratory practices
Convenience:
Practice:No food or drink allowed in the laboratory
Problem: No lunch room
Result:Food stored and consumed in the laboratory
Assumed Risk: Contamination, risk of infection
Inventory:
Practice: Update the inventory at the end of the day
Problem: It’s the end of the day, people are tired
Result:Out to date or incorrect inventory
Assumed Risk: Theft, misuse, loss, confusion
Supporting good lab practices
Positive reinforcement
 Providing solutions to the violations. For example, a lunch table outside the laboratory so
no food is brought or consumed in the laboratory
 Rewards for periods of no violations
 Training/sensitization
Negative reinforcement
 Fines, punitive actions

6.5 Laboratory equipment calibration and validation
Instrument calibration is one of the primary processes used to maintain instrument accuracy.
Calibration is the process of configuring an instrument to provide a result for a sample within an
acceptable range. Calibration refers to the act of evaluating and adjusting the precision and
accuracy of measurement equipment. Instrument calibration is intended to eliminate or reduce
bias in an instrument's readings over a range for all continuous values.
Equipment validation refers to confirm and assure functioning of the equipment as per
required standard. In biosafety and biosecurity perspective validation is highly desirable to
mitigate biorisk. It enables the laboratory managers/supervisors to ensure quality results and
calibrate the equipment if needed within time both for accuracy and biorisk mitigation.
“The organization ( laboratory/department/university) shall establish and maintain
documented procedures to ensure equipment and elements of the physical plant that may impact
on biorisk be identified, purchased, maintained, calibrated, certified or validated in a manner
consistent with the intent and requirements of the biorisk management programme.”
11

42. To validate an instrument, the following points should be considered:
 Identifying and recording validation requirements at time of purchase/acquisition

 Identifying the standards/tests that will be used to ensure the equipment is correctly
validated

 Creating a documented and up-to-date validation register for all applicable equipment

 Ensuring validation is scheduled and conducted in the line with manufacturer’s
requirements and / or other specified intervals as identified by risk assessment

 Ensuring competent and independent validation companies are used for the validation
process.

44. For further study
i. www.who.int/tdr/publications/documents/glp-trainee.pdf
ii. www.who.int/tdr/publications/documents/glp-handbook.pdf
iii. http://www.bcshguidelines.com/documents/Guidelines_for_validation_bcsh_130820
10.pdf


45. Chapter 06:
Personal Protective Equipment
Contents:
 Introduction to personal protective equipment
 Proper use of personal protective equipment
 Donning and doffing of personal protective equipment

Introduction to personal protective equipment (PPE)
Personal protective equipment (PPE) refers to the equipment worn to minimize exposure to a
variety of hazards (bacteria, virus, fungus, toxins, sharp edges, chemicals, flying sparks, falling
objects). Examples of personal protective equipment are gloves, respirators, surgical mask, and
apron, shove covers, goggles and head covers etc.
Types of PPE:
Personal protective equipment mainly includes instruments to cover the exposed parts of
body like hands, face etc and to avoid contact of disease or infections through natural routes of
exposure like mouth, nose, eyes etc. Proper selection of personal protective equipment is based
on nature of hazard, level of exposure to hazard and likelihood and consequences of potential
hazard.
The following are some important personal protective equipment to be used while working in
a veterinary laboratory or handling animals for examination or sample collection in field or
veterinary facility.
Gloves:
 Gloves should be worn when handling infectious materials, chemicals, Glassware, sharp
edge objects, corrosive material and hot objects or animal.
 Gloves must not be slippery and easy to put on and take off.
 Must be made of cotton, nylon, leather or latex as per standard requirement.
 Use accurate kind and size of gloves that fit properly.
 Wash reusable gloves appropriately before removing them.

2. Lab Coat:
A laboratory coat is a knee-length overcoat worn by professionals working in the laboratory
to give maximum protection to the laboratory worker The lab coat must be able to absorb or
deflect splashes or spills so that harmful materials are kept off the skin.
 Lab coats can serve a number of purposes – protection from chemical
Splash, fire resistance or clothing protection.
 The lab coat should offer some protection against flash and fire so that

even if the coat starts to burn, person should escape without injury or
with less serious Injuries.

46. 3. Safety glasses or goggles:
Safety glasses or goggles must be worn when conducting laboratory work with the
possibility for splashing or spraying biological materials
 Face shield in combination with eye protection should be worn when working with
highly infectious agents.
 Ensure that safety glasses fit properly.

4. N95 Respirator:
N95 respirator filters out particulates and liquid droplets in small particle size, therefore
providing protection from inhaling aerosols and microorganisms that are airborne.
Using the appropriate respiratory protective equipment is important for the securing an
adequate protection from biological hazards.
 Ensure that the respirator offers a good fit for the user otherwise user or lab
Workers may not be adequately protected.
 Use appropriate respirator for a biological agent or procedure.
 Surgical mask should not be used where respirator is needed.

5. Surgical Mask:
Surgical mask consist of three layers of non-woven fabrics and provides a barrier, against
large respiratory droplets.
 Before removing the mask from its container and handling it, make sure to clean hands
with soap and water.
 Inspect the mask for tears or holes. Properly discard the disposable mask after use

Use of Personal Protective Equipment:
PPE is an important means of preventing work injuries. It is the best approach to maintain a safe
work environment and eliminate potential hazards.
It is necessary to ensure that the equipment chosen is both reliable and effective, properly used
maintained and the user knows about the donning and doffing methods.
The following key areas need to be considered while using personal protective equipment;
 Proper Selection:
First understand the nature and degree of the potential hazards and then select appropriate
PPE that meets the relevant standards. PPE must meet the demands of the work environment and
should be as comfortable and easy to use as possible.

47.  Correct Use:
Laboratory workers should understand the correct use of the PPE like Donning (putting on
PPE) and doffing (removing PPE).
 PPE fit testing
The principles behind face fit testing are that people, as well as respirators, come in different
shapes and sizes. Therefore choosing a respirator is important if it is to provide the correct
protection for the hazard but critically that it fits the wearer so it gives the optimum protection. If
there is an unsatisfactory seal between the wearer and the hood, or face mask, then it is possible
that leakage could occur and contaminant can enter the breathing zone.
There are two fit testing methods, these are known as: quantitative and qualitative tests.
Qualitative tests rely on the wearer’s subjective assessment of face seal leakage. These methods,
during a set of test exercises, use the wearer’s sense of smell or taste to detect face seal leakage
of a test agent.
Quantitative tests are subject to problems with sensitivity, lack of objectiveness and inability
to provide a numerical result.
The recommended fit testing exercises for both Qualitative and Quantitative tests are:
 Normal Breathing
 Deep Breathing
 Head side to side
 Head up and down
 Talking
 Bending
 Normal Breathing

 Proper Maintenance:
Laboratory workers must clean and dry the reusable PPE after use and properly stored and
regularly inspect it. If the laboratory workers identify any damage to PPE, immediately report it
to the supervisor so that it can be replaced timely. Laboratory workers and students should avoid
taking and cleaning their contaminated laboratory coats at their residence.
 Disposal of personal protective equipment:
The disposable contaminated personal protective equipment like gloves, surgical masks,
shove covers and head covers should be disposed as per standard procedures.
Donning and doffing of personal protective equipment
Donning and doffing of personal protective equipment in proper sequence or order is highly
required to achieve maximum protection.

48. Donning sequence
1- Lab coat
 Fully body cover from neck to knees, arms to end of wrists and wrap around the back.
 Fasten in back of neck and waist.

2- Mask or respirator
 Secure ties or elastic bands at middle of head and neck
 Fit flexible band to nose bridge
 Fit comfortably to face and below chin
 Fit-check respirator
3- Goggles or face shield
 Place over face and eyes and adjust to fit

4- Gloves
 Extend to cover wrist over lab coat cuffs.
Doffing (removing) sequence
1. Gloves
 Outside of gloves is contaminated so grasp outside of glove with opposite gloved hand
and peel off.
 Hold removed glove in gloved hand
 Slide fingers of ungloved hand under remaining glove at wrist
 Peel glove off over first glove
 Discard gloves in waste container

2. Goggles or face shield
 To remove, handle by head band or ear pieces as outside of goggles or face shield is
contaminated
 Place in designated receptacle for reprocessing or in waste container

3. Lab coat
 Coat front and sleeves are contaminated so unfasten ties and pull away from neck and
shoulders, touching inside of gown only
 Turn gown inside out
 Fold or roll into a bundle and discard

49. 4. Mask or respirator
 Front of mask/respirator is contaminated — DO NOT TOUCH
 Grasp bottom, then top ties or elastics and remove
 Discard in waste container

51. For further reading
1. http://www.cdc.gov/HAI/pdfs/ppe/ppeposter148.pdf
2. http://www.nebraskamed.com/app_files/pdf/biocontainment/ebolappechecklist.pd f
3. http://www.cdc.gov/vhf/ebola/healthcare-us/ppe/guidance.html

52. Chapter 07
Biosecurity
Contents:
 Introduction to biosecurity
 Physical security
 Personal management
 Material control and accountability
 Transportation security
 Information security

Introduction to biosecurity:
The term biosecurity is more complex as it can have different meanings in different contexts.
According to the relevant WHO guidance, the phrase has evolved simultaneously in various
processes and is used differently in each.
The term biosecurity does not seem to have a standardized meaning across human, animal
and plant health sectors. In veterinary and agricultural fields, the term has come to denote
protecting biological resources from foreign or invasive species. 10
Or
Biosecurity describes the protection, control and accountability of valuable biological material.11
Or
At the Biological Weapons Convention (BWC) meetings in 2003, one delegate used this
simple formulation to assist participants differentiate between biosafety and biosecurity issues:
“Biosafety protects people from germs – biosecurity protects germs from people”.
Biosecurity refers to the principles, technologies and practices that are implemented to secure
pathogens, toxins and sensitive technologies from unauthorized access, loss, theft, misuse,
diversion or intentional release.
Biosecurity covers a broad spectrum of potential risks and threats ranging from criminal
activities, such as sabotage and isolated acts of aggression, to bioterrorism and spying. The term
‘biosecurity’ has been used in discussions of the risks associated with dual uses of the life
sciences, where legitimate research may have malicious applications and implications beyond its
intended use. The term is also used to describe environmental risks in the areas of agriculture and
food safety. Discussions of the effect on biodiversity of the introduction and release of
genetically modified organisms or foreign invasive species use the term as well.
10
Biorisk management Laboratory biosecurity guidance September 2006 WHO/CDS/EPR/2006.6
11
Office of International des Epizootics (OIE)

53. History:
Misuse of valuable biological material:
Case1. In September and October 2001, several cases of anthrax broke out in the United States
anthrax attacks, apparently caused deliberately. Letters laced with infectious anthrax were
concurrently delivered to news media offices and the U.S Congress, alongside an ambiguously
related case in Chile. The letters killed 5 persons12
.
Case2. In 1996 a laboratory technician at St Paul’s Medical Center in Dallas, Texas, USA, laced
pastries with Shigella dysenteriae type 2, causing 12 of her co-workers to become ill.
Premeditation was apparent: she had sent an email to her colleagues saying that pastries were
available in the coffee room.13
Case3. In 1970 a postgraduate student of Parasitology at the Macdonald campus of McGill
University near Montreal, Canada, contaminated his room-mates’ food with stolen Ascaris suum
eggs. Two of the four room-mates developed acute respiratory failure before parasitic worms
was identified as the cause. Prior to the incident, there had been numerous disputes about the rent
for the apartment where the student lived with his room-mates.14
Objective:
The objective of laboratory biosecurity is to safeguard materials, employees, information and
other laboratory assets. This is true regardless of whether the laboratory’s activities relate to
research, diagnostics, the production of pharmaceuticals or to other life science areas.
Biosecurity addresses laboratory risks and threats that fall outside the domain of accidental
exposure or release.
A – To reduce the risk of theft and fraud.
B – To reduce the risk of scientific misconduct
Key Pillars of biosecurity:
Implementing biosecurity encompasses many aspects of laboratory activities and resources.
It aims to ensure the integrity and security of all pathogens, toxins and sensitive information.
Biosecurity has a number of key components.
 Physical security
 Personal Management
 Information security
 Transport security
 Material control and accountability


12
http://edition.cnn.com/2008/CRIME/08/06/anthrax.case/index.html
13
http://www.readcube.com/articles/10.1002%2F0471686786.ebd0154.pub2
14
Carus, S. W., ‘Bioterrorism and biocrimes: the illicit use of biological agents since 1900’, Working paper, Center for
Counter proliferation Research, National Defence University, Washington, DC, Feb. 2001, pp. 43–45, 61

54. Physical security:
Physical biosecurity, comprised of engineering, structural and security personnel elements, is
intended to select, control and document, access to laboratories and to the materials they contain,
and to limit improper removal of VBM and equipment. Access controls are used to limit access
to restricted areas to individuals who don’t have proper authorization and to keep track of traffic
in and out of these areas. Physical biosecurity measures may become more strict and more costly
as the value of the assets increases and as the location of the materials to be protected is
approached.
Example:
 Security guard/walk through gates
 Solid doors with lock
 Bars on windows
 Identification cards/PIN (Personal Identification Number)
 Cameras
 Boundary walls and fences
 Barriers

Personnel management:
Personnel management procedures should define the roles, responsibilities and authorities of
laboratory personnel who need to handle, use, store, transfer and/or transport VBM, and the
manner in which the organization ensures that individuals are appropriate for the positions they
hold. These procedures should clearly describe and document the training, experience,
competency and suitability requirements for individuals who have access to VBM, ensuring that
members of the workforce have appropriate personal and technical qualifications and skills.
Documented procedures for the recruitment of personnel should be clearly established and
followed. The professional and bioethical eligibility and suitability for working with VBM of all
personnel who have regular authorized access to sensitive materials is also central to effective
laboratory biosecurity risk management.
A mechanism should be developed to ensure that the integrity of the facility will not be
compromised through the absence of key individuals. Such a mechanism should include
succession planning for management, scientific, technical and administrative personnel to ensure
that critical knowledge of safe and secure operation of the facility does not reside with a single
individual, in the event of his/her unavailability or departure. Provisions describing personnel
management should also address procedures and training/instructions for visitors, contractors,
subcontractors, suppliers, cleaning and maintenance staff.
Information security:
Information security establishes wise policies for handling sensitive details on VBM.
Examples of sensitive information may include laboratory security plans and inventories and
storage locations of VBM. Information security should ensure that the required and appropriate
level of confidentiality is preserved by the system that is used to acquire, store, manipulate and
manage information. It is important to establish practical and realistic steps that can be taken to
safeguard and track VBM. A comprehensive documentation and description of VBM represents
the caretaker role of the current laboratory managers to accurately pass on the historical archive

55. of VBM. Some of the information may be confidential but should be available for use by future
generations. Such documentation may also prove useful to help discharge a facility from possible
allegations. It is also important to document the existence, location and access to the information
for future interests, as security concerns change over time. The higher the level of risk associated
with the VBM the institution holds, the greater protection the information associated with the
security system will require. Overdoing or exaggerating the sensitivity or level of suspicion can
have unintended negative repercussions. Laboratory management and relevant authorities should
develop appropriate policies that govern the marking and handling of information and how that
information is gathered, maintained, distributed, documented, accessed, shared and stored within
the facility and with appropriate counterparts.
Transport security:
Transport security refers to provide a measure of security during the movement of biological
materials outside of the access-controlled areas in which they are kept until they arrive at their
destination. Transport security applies to biological materials within a single institution and
between institutions.
1. Internal material transport security includes reasonable documentation, accountability
and control over valuable biological material moving between secured areas of a facility
as well as internal delivery associated with shipping and receiving processes.
2. External transport security should ensure appropriate authorization and communication
between facilities before, during and after external transport, which may involve the
commercial transportation system..
Transfer of materials:
Many countries request to file import and export permits for biological materials before the
transfer of such specimens is authorized. These procedures allow for registering and tracking of
materials entering or leaving a country and they are particularly important in the case of alien or
dangerous pathogens.
Example:
1. Live Bacillus anthracis shipped by mistake
In May 2004 live Bacillus anthracis was shipped from the Southern Research Institute
in Frederick, Maryland, USA, to the Oakland Children’s Hospital and Research Center in
Oakland,
California, USA. The sender had verified that the samples were not viable. However, the
death of 49 mice that were infected using the Bacillus anthracis samples revealed the
viability of the samples.15
15
Vesely, R., ‘Anthrax incident spurs concern’, Oakland Tribune, 12 June 2004

56. 2. British journalists order a sequence of modified smallpox DNA
On 14 June 2006, The Guardian reported that journalists had successfully ordered online and
received a plastic vial containing the 78bp sequence of DNA coding for the smallpox virus coat
protein modified with three mutations from VH Bio Ltd. The vial, which arrived in an A5-size
Jiffy bag, cost £33. The company was unaware that the sequence belonged to the smallpox
genome.16
Material control and accountability:
Proper inventory practices are essential for effective material control and must cover
pathogens and toxins from the time that they arrive at a facility or laboratory to their final
destruction or shipping. Proper inventory practices must include the confirmation of receipt by
the designated party.. The employees who have access to and work with biological materials
must be responsible for basic inventory tasks because they are familiar with the type and amount
of biological materials present and their location and state.
Rules and procedures for inventory control must be developed by laboratory managers or
principal investigators, together with management and biosecurity officer.
Inventories of pathogens and toxins should include relevant information such as:
• Types of material (name, strain, serotype, taxonomy etc.);
• Forms of material (solution or pellet, freeze dried, paraffin embedded etc.);
• Quantities of material (number of vials, amounts of liquid, post-experiment quantities);
• Locations of material (in short- or long-term storage or in use);
• Contact or responsible employee;
• Employees who have access to the materials;
• Confirmation, date and method of disposal, destruction or inactivation of material
• Dates of transfer of material (delivery and departure) and end-user or recipient receipts.
• A general material list for the whole facility can be maintained, coordinated and managed
by the appointed biosafety and biosecurity officer.
16
Randerson, J., ‘Revealed: the lax laws that could allow assembly of deadly virus DNA’, The Guardian, 14 June
2006.

57. For more detail please visit:
1. http://www.who.int/csr/resources/publications/biosafety/WHO_CDS_EPR_2006_6.pdf
2. http://aseanregionalforum.asean.org/files/Archive/6%20ARF%5BMr_%20Stuart%20Bla
cksell%5D%20-%20BW.pdf
3. http://www.oie.int/fileadmin/Home/fr/Health_standards/tahm/1.01.03_BIOSAFETY.pdf

58. Chapter 08:
Incident Management
Contents:
 Scope of incident management
 Incident response system
 Incident report

Incident Management:
Incident: An instance of something happening; an event or occurrence.
Management: The process of dealing with or controlling resources and people in an efficient
way.
Incident vs. Emergency:
 An incident is an event that is likely to have adverse consequences. An emergency is
an incident that requires an immediate response.
Example of Incident:
• Energy failure
• Breakage of a container having pathogen
• Spills
• Cuts and wounds
• Pricks
• Chemical burn, fire and short circuit
• Bio Safety Cabinet suddenly stop working
• Laboratory controlled air system stop working
Incident Response System:
Every Incident Response System must be able to alert to an incident and mobilize a
response.
Five components of Incident Response System:
1. Planning
2. Preparation
3. Alert
4. Assessment
5. Mobilization

59. Model for an Effective Incident Response System:
Planning Preparation
Incident
Alert Assessment Mobilization
Incident Response System:
• The most effective incident response systems helps in effective planning and preparation
for potential incidents, alert and assess actual incidents and quickly mount an effective
response if an incident occurs.
• Management has the authority to make medium and long-term decisions and
Allocate appropriate resources towards an incident management system.
1. Planning
Planning is the development of mechanisms or procedures, in advance, to achieve a
particular goal.
E.g. Written policies and procedures for incident management (SOPs)
In the case of incident response, planning is the process whereby a potential incident is
considered and evaluated and resources are assigned in order to generate a response that will
appropriately mitigate any adverse effects.
2. Preparation:
• Preparation derives directly from planning. It is the act of putting into effect an
institution’s plans prior to an incident, in order to be in a position to better handle that
incident when it does occur.
• The Preparation process includes training of personnel (Drills), acquisition of equipment
(First aid equipment, fire extinguisher etc), storing of supplies and physical modifications
to equipment and buildings when possible and desirable.

60. Model for Incident Management:
Planning and Preparation
Incident Drill
Alert, Assessment, and Mobilization
3. Alert:
 Alerting is the process of identifying an incident as it is occurring, or after it has occurred
and using that information to generate a response. E.g. symbol, alarm, signs.
 An institution’s incident response plan, as developed by management, should address
procedures for alerting appropriate personnel in the event of an incident.
4. Assessment
Assessment is the evaluation of the type and severity of an incident, in order to determine an
appropriate response.
Alerting and assessment provide information as to the existence and nature of a particular
incident.
Some incidents are self-resolving, and by the time an alert is raised and the incident is
assessed, a specific response may not be needed.
5. Mobilization:
Mobilization is the activation of personnel and use of equipment to respond directly to an
incident and speed up its resolution.
Feedback:
Feedback is the use of information gathered from drills or incidents to inform and
improve an Incident Response System.

61. Specifically, feedback informs the planning and preparation process, providing information
to management and other personnel on what procedures were and were not effective in
mitigating the adverse consequences of incidents (or drills).
Outside Responders:
As part of planning and preparation as well as alerting, assessment, and response, an
incident response system must interact with the outside world in order to properly manage
serious incidents. It is often not economical for an institution to develop the capacity to
respond to large, infrequent incidents and emergencies completely on its own. Also,
depending on the laws of the jurisdiction that an institution is located in, there might be
certain kinds of incidents that the institution cannot respond to. Thus, coordination with
police, fire brigades, and emergency medical services are crucial for effective incident
response.
Incident Report:
Incident report is one of the important activities after each incident. It enables the
management to timely respond and extend support in case of any incident and emergency as
well as developing planning for incident management. There should be standard operating
procedure for incident reporting and incident report form in each laboratory or
health/veterinary facility.

63. For further study
3. http://www.safety.uwa.edu.au/incidents-injuries-emergency/procedures/lab
4. https://www.ehs.umass.edu/lab-incidents-and-lab-incident-report-form
5. Laboratory biosafety manual WHO 2006 edition

64. Chapter 09
Waste Management
Contents:
 Introduction to waste management
 Categories and steps in waste management
 Segregation
 Storage
 Treatment of different waste
Introduction to Waste management
Waste is anything that is to be discarded. In laboratories, decontamination of wastes and their
ultimate disposal are closely interrelated. In terms of daily use, few contaminated materials will
require actual removal from the laboratory or destruction. Most glassware, instruments and
laboratory clothing will be reused or recycled. The prime principle is that all infectious materials
should be decontaminated, autoclaved or incinerated within the laboratory.
19
Biological Waste Management
Infectious waste, pathological waste, bio-contaminated sharps and other waste from laboratories
and clinical areas are considered as bio-hazardous wastes which needs special handling and
disposal. Infectious wastes need pre-treatment before they are disposed of.
Biohazardous Waste:
Waste by their nature and quality may be potentially harmful to health of human, animal and
environment.
Waste management requires:
 Characterizing waste
 Segregation hazardous waste
 Collection
 Packaging and labeling
 Storage of waste
 Treatment of waste
Hazardous Waste
Any waste that directly or indirectly poses a threat to human/animal health or to the environment
by introducing one or more of the following risks:
 Pathogens, parasites or their vectors
 Explosion or fire
 Chemical instability, reactions or corrosion
 Acute or chronic toxicity
 Toxicity or damage to the ecosystems or natural resources
 Accumulation in the biological food chain, persistence in the environment or multiple
effects.

65. Types of laboratory waste
 Solid waste (Sharp and Non Sharp)
 Liquid waste
 Pathological waste

Solid waste (Non Sharp)
 Gloves and other disposable PPE contaminated with specimen or culture material.
 Plastic ware such as pipettes or pipette tips, culture plates, specimen vials, etc. that are
contaminated with biological specimens, bacterial and cell culture material or nucleic
acids.
 Towels and bench paper that are biologically contaminated.
 All culture or sample containers that are contaminated with biological materials
 Tubes of blood (Note: glass blood vials that could break easily upon disposal should be
segregated as sharps waste.).
Collection and storage:
Collect waste in a solid-walled, leak-proof container lined with an autoclaveable biohazard
bag. The container needs to have a lid and be marked with the biohazard symbol. The container
needs to be closed when not actively in use. If the container has no lid, the bag needs to be
securely closed and transferred to the floor container at the end of the day. Collect serological
pipettes separately in order to prevent bag puncture during the waste handling process.
Treatment and disposal:
Bags should be securely closed and transferred to a leak-
proof secondary container in the designated bio-hazardous waste
pickup point in the lab. (Heavy bags should be double-bagged to
further prevent leakage during the handling process).
These waste bags are autoclaved or incinerated for final
treatment by the vendor or by the lab at site.
2. Solid waste (Sharps):
It includes items that can puncture or tear autoclave bags e.g.
scalpel blades, hypodermic needles, syringes (with or without
attached needles) and needles with attached tubing, disposable
pipettes, razor blades, blood vials, test tubes, pipette tips, broken
plastic culture dishes, glass culture dishes and other types of broken
and unbroken glass waste (including microscope slides and cover
slips) that may have been in contact with infectious material.
Collection and storage:
These sharps should be put into sharps container immediately after
use. Sharps containers must be biohazard-marked, solid-walled,
puncture-proof containers that are leak-proof on the sides and bottom.
Sharps

66. container lids have a restricted access opening to prevent devices from being accessed once
inside the container. Assure that the lid is properly and completely installed before using it for
sharps collection.
Treatment and disposal:
Permanently close the container when it is three quarters full or when items do not freely fall
into the container, regardless of the fullness level. Place filled containers in the designated bio-
hazardous waste pickup point. Container for sharps must be autoclaved using a validated waste
treatment cycle or incinerated for final treatment.
3. Liquid waste:
Blood and blood product, semen, vaginal secretions, cerebrospinal fluid, synovial fluid,
peritoneal fluid and pericardial fluid and culture broth.
Collection and storage:
Vacuum flasks and liquid “pour-off” containers should be charged with disinfectant prior to
use to help prevent growth of contamination in the flask during the holding period. Collection
vessels need to be labeled with the biohazard label and name of the disinfectant. Non-breakable
vessels should be used whenever possible.
Vacuum flasks should be stored in a non-breakable and leak-proof secondary container when
not maintained inside a biosafety cabinet (BSC). Vacuum flasks must also be equipped with an
overflow flask and/or HEPA filter on the line to protect vacuum lines in the event of a flask
malfunction. Flasks should be discharged and cleaned weekly, or when they are half-full,
whichever comes first, to prevent overflow and prevent growth of contaminants.
Treatment of liquid waste:
All liquid waste should be treated prior to disposal. The aim is that the waste does not
contaminate the surrounding areas. To ensure that the contamination is not spread/leaked to the
environment all waste including liquid waste should be treated prior to disposal. There are
several ways/means for decontamination of waste, which are enlisted as under: -
 Disinfection
 Sterilization
 Heat (Dry and Moisture). Autoclave and incinerator
 Ultra Violet rays, i.e Biosafety cabinet
 Chemicals. Formaldehyde, Phenyl, Bleach
Disposal:
Disposal of waste is the process by which the treated waste is done away with. The liquid
waste may be disposed of by washing away into sink, by dumping in designated garbage or by
handing over to authorized contractors.
4. Pathological waste
This includes all unfixed human/animal tissues, organs, body parts and carcasses that have
been exposed naturally or experimentally challenged with infectious agents.

67. Storage and Disposal:
This type of waste must be double-bagged in biohazard bags that bear a biohazard symbol.
Bags must be stored in a manner that will minimize the potential for release of fluids during the
storage and handling process.. These items must be incinerated or treated by alkaline digester
(not autoclaved).
General guideline:
Segregation:
 Pathological waste must be stored in a secure environment at all times.
 Whenever possible, pathological waste must not be mixed with chemical, radioactive or
other laboratory trash.
 The various types of pathological waste should be segregated from each other
 Fluid waste should be pathological contained separately from solid waste.
Containment and Labeling:
Containers for pathological waste must be appropriate for its contents. There are several
different kinds of containers and bags available for the containment and disposal of Pathological
waste. All containers for pathological waste must display the biohazard symbol.
Anatomical/Animal Waste:
Place waste in a red biohazard bag or a black garbage bag if the waste will immediately be
stored in a barrel lined with red biohazard bags. Barrels are approved heavy duty rigid cardboard.
Label: Each bag must be tagged with a completed yellow incineration (necropsy) tag.
Biological Laboratory Waste:
Solid: Collect waste in a red or orange autoclaveable biohazard bag and label
Liquid: Use leak-proof containers which are able to withstand thermal or chemical treatment
The following are some guidelines to remember when packaging waste:
 Double bag if necessary to prevent perforations
 Add absorbent material if the possibility of large volumes of liquid exists.
 Ensure the bags are well sealed
 Do not overfill the containers/bags
 If the outside of the bag is contaminated, use double bag.
 Secondary containment should also be labeled with the biohazard symbol.
Records:
As biomedical waste is potentially hazardous, it is important to document the nature of the
waste. Labels attached to waste bags or sharps containers should detail the contents. Each waste
generator should have a system to track the waste.