Safety Manual

Of
XXXXX
Issue No: xx
Issue Date: xx/xx/xxxx
Safety Manual
 Safety Manual

 Revised / Updated procedures

Sr. No Procedures Page No.

1 Introduction Page no 06
2 Eating /Mouth Pipetting Page no 19
3 Hepatitis vaccination declination Page no 20
4 H1N1 Vaccination distribution Page no 21
5 Tetanus vaccination Page no 21
6 Covid-19 Vaccination Page no 21
7 Safety officer details Page no 30
8 Emergency eyewash Page no 40
9 Covid-19 Safety Measures Page no 67
10 Liquid waste disposal Page no 42
Guidelines for Surveillance handling microorganisms
11 Page no 48
at Biosafety level
12 CO2 Fire extinguisher Page no 34
13 ETP Page no 43
14 Biohazard disposal Page no 43
15 Sharp containers Page no 45
16 Autoclave procedure Page no 45
Guidelines for the surveillance of laboratory
17 Page no 48
workers handling microorganisms at Biosafety Level
3
18 First aid box & Spill kit Page no 42
19 ALL BL3 Special Practices Page no 57
 Safety Manual

Contents

Introduction ......................................................................................................................................................6

1. Accidents in the laboratory.............................................................................................................6

a) Unsafe premises..............................................................................................................................6
b) Naked flames....................................................................................................................................7
c) Microbial hazards............................................................................................................................8
d) Chemical hazards............................................................................................................................9
e) Glassware hazards.......................................................................................................................10
f) Equipment hazards......................................................................................................................10
g) Explosions.......................................................................................................................................11
h) Infestation by ants, rodents, cockroaches ...........................................................................11
i) Unreliable water supply.............................................................................................................12

2. Occupational Injuries ......................................................................................................................12

3. Emergency preparedness plans..........................................................................................................13
4. Evacuation Plan................................................................................................................................15
5. Blood borne pathogens....................................................................................................................5
a) PPE Provision and Usage...........................................................................................................15
b) Hand Hygiene................................................................................................................................16
c) Manual Manipulation of needles.............................................................................................19
d) Eating /Mouth Pipetting.............................................................................................................19
e) Specimen Transport Procedure........................................................................................................19
f) Spill Handling.................................................................................................................................19
g) Hepatitis B Vaccinations............................................................................................................20
h) H1N1 Vaccination.........................................................................................................................20
i) Covid-19 Vaccination...................................................................................................................21
j) Tetanus vaccination......................................................................................................................21
k) Viral Exposure...............................................................................................................................22
l) TB Exposure.....................................................................................................................................31
m) Sterilizing Device Monitoring..........................................................................................................32
6. Fire Prevention and Protection...................................................................................................32
a) Fire Separation..............................................................................................................................33
b) Fire Exit.....................................................................................................................................................33
c) Annual Fire Drill............................................................................................................................33
d) Fire detection/Alarm...................................................................................................................33
e) Fire Extinguishers..................................................................................................................................34

7. Electric Safety...................................................................................................................................34
8. Chemical Safety................................................................................................................................34
a) Chemical Hygiene plan...............................................................................................................34
b) MSDS.................................................................................................................................................35
c) Chemical Precautionary Labels...............................................................................................35
d) Flammable, Acid Base & Volatile solvent Storage....................................................................35
 Safety Manual
9. Environmental Safety.....................................................................................................................36
a) Laboratory Ergonomics Program.....................................................................................................36
b) Excessive Noise.............................................................................................................................40
c) Emergency Eyewash...................................................................................................................40
10. Other Hazards...............................................................................................................................40
a) UV Light Exposure........................................................................................................................40
b) Latex Allergy..................................................................................................................................40
c) Dry Ice hazards and Safe handling ........................................................................................41
d) First aid box & Spill kit...............................................................................................................42

11. Waste Disposal.......................................................................................................................................41

a) Liquid Waste Disposal..........................................................................................................................41
b) Biohazard Disposal Containers.........................................................................................................42
c) Sharp Disposal...............................................................................................................................42
d) Autoclave procedure...................................................................................................................46
12. Microbiology safety...............................................................................................................................44
a) Hazardous Procedure...........................................................................................................................44
b) Laboratory Biosafety Levels..............................................................................................................44
c) Preventive Measures against Laboratory Infections ..........................................................45
d) Biosafety Precautions in TB lab...............................................................................................................48
13. Bioterrorism plan.........................................................................................................................54
14. Covid-19 Safety measures........................................................................................................61
a) Social Distancing............................................................................................................................62
b) Hand Hygiene.................................................................................................................................62
c) PPE and Usage................................................................................................................................62
d) Decontamination............................................................................................................................65
e) Other Safety measures................................................................................................................68
 Safety Manual

I n t r o d u c t io n
The laboratory environment can be a hazardous place to work. Laboratory staffs
are exposed to numerous potential hazards including chemical, biological, physical
and radioactive hazards, as well as musculoskeletal stresses.
This laboratory Safety manual is compiled to be used as a binding document for all
personnel working to ensure safe work conduct and practices in XXXXX.
Procedures and Rules within this Manual are formulated for three reasons:
 To avoid health risks and accidents for our personnel.
 To be in a position to act appropriately in case of emergencies.
 To minimize the environmental burden and risks caused by our work.

1. Accidents in the laboratory

 Injury, ill health, and disablement of staff, patients and others.
 Work being disrupted with possible discontinuity of laboratory services.
 Loss of valuable laboratory equipment, supplies, and records.
 Loss of or contamination of specimens with serious consequences for patients.
 Damage to the laboratory, adjacent departments, and to the environment.
If such accidents are to be avoided, a relevant, workable, and affordable
laboratory health and safety program is essential.
Common hazards in laboratory that require assessment and management are:

a) Unsafe premises
Burns & inhalation of smoke during a fire:
 When emergency exit routes from the laboratory are blocked by
equipment, storage boxes, etc.
 When, in a subdivided laboratory, there is only a single exit and the staff
becomes trapped in one section.

Staff are injured by falling on a slippery or damaged floor or from broken glass
on the floor:
 When the floor is not cleaned properly after spillages or glassware breakages.
 When wax or other slippery cleaning substance is applied to the floor.
 When damaged areas of the floor are covered with matting.

Risk of infection to staff & others:

 When there is no separate hand basin with a reliable water supply for
hand washing.
 When no separate wash-room is provided for staff and food and drink are
consumed in the laboratory.
 When laboratory staff do not leave their protective clothing in the
laboratory when leaving the workplace or when the clothing is not
laundered frequently enough.
 Safety Manual

 When bench surfaces are not disinfected or cleaned properly each day.
 When the work area is not separated from the areas where outpatients
are received and blood samples collected.
 When the laboratory has no safe systems for decontaminating infective
materials, disposing of waste and washing reusable laboratory ware.

Injury from chemicals:

 When chemicals with irritating fumes are used in a laboratory with
inadequate ventilation.
 When hazardous chemicals are stored on high shelves or on the floor
under benches.

Injury from equipment:

 When electrical equipment has faulty earthing or insufficient ventilation.
 When adaptors or extension leads are used because there are insufficient
electric wall points.
 When the laboratory has no preventive maintenance schedules and
equipment is not inspected regularly for defective insulation, corrosion,
and loose connections.

Unsafe Laboratory Premise-Prevention

 Laboratory premise is structurally sound and in good repair with a reliable
water supply and safe plumbing and waste disposal system.
 Adequate floor and bench space and storage areas are available.
 Floor surface is non-slip, impermeable to liquids, and resistant to those
chemicals used in the laboratory.
 Walls are smooth, free from cracks, impermeable to liquids, and painted with
washable colour paint.
 Adequate ventilation is supplied by wall vents and windows that can be
opened.
 Phlebotomy area is away from the testing area of the laboratory.
 Bench surface area is without cracks, impervious, washable, and resistant to
the disinfectants and chemicals used in the laboratory.
 A suitable storage facility for storage of chemicals is provided.
 A staff room is separate from the work area where refreshments can be taken
and personal food and other belongings stored safely.
 Safe electric supply with sufficient wall electric points to avoid the use of
adaptors and extension leads. The electrician will check all points every 6
months and put the all O.K. sign on the point.
 Fire extinguishers sited at accessible points.
 Containers for decontamination of infected material, discarding of needles,
syringes, lancets etc. are labelled with a warning symbol such as a red
triangle.

b) Naked flames
Injury from fire caused by lighted Bunsen burners, spirit burners, tapers,
matches, alcohol swabs, ring burners, stoves:
 Safety Manual

 When a lighted burner is paced in sunlight, making the flame difficult to see.
 When a burner, match, or taper is lit too close to a flammable chemical.
 When a ring burner or stove is positioned too close to where flammable
chemicals are used or stored.

Naked flames-Prevention
1) When lighting and using a Bunsen burner always check that there is no
flammable chemical or reagent nearby.
2) Be particularly careful when heating carbol fuchsin stain on slides in Ziehl-
Neelsen technique. Use only a small lighted swab and extinguish it
immediately after use. Keep the flame well away from acid alcohol,
acetone, methanol, methylated spirits, ether stains such as Giemsa and
Leishman, and other flammable reagents.

c) Microbial hazards
Pathogens area accidentally ingested:
 From contaminated fingers when personal hygiene is neglected.
 When hands are not washed after handling specimens or cultures.
 Mouth pipetting is absolutely forbidden.

Pathogens are accidentally inoculated:

 Through needle stick injuries caused by resheathing needles after
collecting blood or careless handling of needles and lancets.
 Through open, uncovered skin wounds.
 Through injury from broken contaminated glassware.

Pathogens are accidentally inhaled in airborne droplets (aerosols):

 When snap closing specimen containers.
 When vigorously dispensing or pouring infectious fluids.
 When sucking up and blowing out fluids from pipettes.
 When during centrifugation a container breaks and the lid is opened
before the aerosols have settled.
 When infectious material is spilled following the dropping or knocking over of
a specimen container or culture.

Microbial hazards-Prevention
Good technique and the practice of personal hygiene are the most
important ways of reducing contact with infectious material and
preventing laboratory related infections.
 Washing of hands and arms with soap and water before and after
attending outpatients, visiting patients, after handling specimens and infected
material, when leaving the laboratory, and at the end of the day’s work.
 Covering any cuts, insect bites, open sores, or wounds on the hands or other
exposed parts of the body with a water-proof adhesive dressing.
 Wearing closed shoes and not walking barefoot.
 Not eating, drinking, chewing gum, smoking, or applying cosmetics in any
part of the laboratory and not sitting on laboratory work benches.
 Safety Manual

 Food and drinks should never be stored in a laboratory refrigerator.

 Not licking gummed labels or placing pens, pencils, or other articles near the
mouth, eyes, or in hair.
 Avoid wearing jewellery in the working area, particularly pendant
necklaces and bracelets.
 When needing to pour infectious fluids, for e.g. in disposing of
supernatants that cannot easily be pipette, the fluid should be poured
carefully down the side of a funnel.
 Specimens, culture tubes and bottles should always be placed in racks so
that they cannot fall over and spill their contents.
 Mouth pipetting is prohibited due to the high risk of infection and injury to
laboratory staff.

d) Chemical hazards
Toxic or harmful chemicals causing serious ill health, injury, or irritation:
 When toxic or harmful chemicals are swallowed by being mouth pipetted.
 When fumes from irritant chemicals are inhaled in poorly ventilated areas
of the laboratory.
 When no protective goggles or gloves are worn and harmful chemicals
enter the eye or come in contact with the skin.

Flammable chemicals causing fire:

 When flammable chemicals are used or stored near a naked flame.
 When a lighted swab is used to heat stain in the Ziehl-Neelsen method
and ignites nearby flammable chemicals.
 When the neck of a bottle containing a flammable chemical is accidentally
flamed.
 When a flammable chemical is spilled near a flame.

Corrosive chemicals causing serious injury and burns:

 When corrosive agents are ingested by being mouth-pipette.
 When strong acids are accidentally knocked from shelves or spilled.
 When intense heat is produced during the dilution or dissolving of a strong
acid or alkali, e.g. adding water to concentrated sulphuric acid.
 When a corrosive chemical comes into contact with the skin, or the eyes
are splashed when opening and pouring a corrosive material.

Chemical hazards-Prevention
 The laboratory is kept well ventilated to prevent any build-up of
flammable gases and vapours.
 Before opening a bottle of flammable liquid, always make sure there is no
open flame within 2 meters such as that from a Bunsen burner or a
lighted candle.
 Ensure stock bottles and dispensing containers of flammable liquids are tightly
closed after use.
 Do not light a match or use a lighted taper near to a flammable chemical.
 Make sure no one smokes in or adjacent to the laboratory.
 Safety Manual

 Place dispensing containers of acetone, acid alcohol, methanol, and

alcoholic Romanowsky stains well away from the rack used to heat stain
on slides in the ZN technique for AFB staining.
 Use trays to hold the containers to prevent a flammable liquid spreading
should a spillage occur.

e) Glassware hazards
Broken glass causing cuts, bleeding, and infection:
 When cleaning damaged slides or cover glasses.
 When using pipettes with broken ends that have not been made smooth in
a flame.
 When picking up pieces of glass following a breakage.
 When glass fragments are left on the floor after a breakage.
 When a waste bin containing broken glass is overfilled.
 When glass and other sharp articles are not separated from other refuse
or the sharps are discarded in containers that can be easily punctured.

Glassware hazards-Prevention
Safe handling of glassware:
 Use appropriate plastic containers for soaking and decontaminating used
glassware. Minimize damage, breakage, and risk of injury.
 Before reuse, inspect glassware, particularly tubes, pipettes and specimen
containers for cracks, broken and chipped ends.
 Never centrifuge cracked tubes or bottles.
 Wear protective gloves when cleaning glassware and avoid overcrowding
drainage and drying racks.
 Laboratory glassware should never be utilized as food or beverage
containers.
 Store glassware safely. Do not leave it in open trays or other places where
it can be easily damaged.
 To avoid spillages and breakages, use racks or trays to hold specimen
containers and other bottles.

Safe management of breakages:

 Wear gloves and use forceps to collect broken glass. A dust pan and small
broom is provided for collecting glass fragments.
 Discard broken glass in a separate puncture resistant sharps container
and dispose of contents safely.
 Treat immediately any glass injury and cover cuts with an appropriate
dressing.

f) Equipment hazards
Electric shock:
 When equipment is not reliably earthed or electrical circuits are faulty.
 When touching live wires in attempting to repair equipment or replace
components, e.g. lamp, without first disconnecting the equipment from
the mains.
 Safety Manual

 When handling electrical equipment with wet hands or standing on a wet

floor.

Fire:
 When cables and electrical equipment overheat due to overloading of
conductors.
 When there is overheating caused by the overuse of adaptors.
 When insulation is inadequate or becomes damaged.
 When thermostats fail and there is no temperature cut-out device to prevent
overheating.
 When electrical sparking or arching causes flammable material to ignite.
 When preventive maintenance is not carried out to check for corrosion, wear,
and loose connections.
 When a battery lead becomes accidentally positioned across the opposite
battery terminal.

Injury from moving parts:

 When a person opens a centrifuge lid and tries to stop the motor manually
(where the equipment does not have a safety device to prevent this).
 When a centrifuge is not balanced, resulting in the buckets and trunnions
spinning off the rotor, particularly when there is corrosion.

Equipment hazards-Prevention
 All personnel must know the location of master switches and circuit
breaker boards. Do not attempt to repair any instrument while it is still
plugged in.
 Plugs or cords that are broken, frayed, or worn should not be used.
 Outlets must not be overloaded.
 All cord and plug-type electrical equipment should have grounded power
cords and plugs. All shocks, including small tingles, must be immediately
investigated.
 Use of extension cords should be discouraged.

g) Explosions
Injury from explosions:
 When incompatible chemicals explode.
 When bottles of fluid explode inside an autoclave.

Explosion hazard-Prevention
The safe use and storage of chemicals are same as above

h) Infestation by ants, rodents, cockroaches

Damaged equipment causing injury:
 When insects enter unmeshed ventilation openings, leading to damaged
components and electrical faults.
 When rodents damage earthing and insulation around cables causing
electric shock and fires.
 Damage to structure and furnishings of the laboratory:
 When ants and rodents damage wooden window frames, bench supports
or shelving.
 When there is no inspection of the laboratory or an infestation is not treated.

Insect & rodent infestation-Prevention

Pest control is done once in three months for ants & cockroaches. If after
a pest control service pests are seen in the laboratory area then, the pest
control contractors are contacted for a re treatment.

i) Unreliable water supply

Contributing to infection:
 When there is insufficient running water for hand washing and the laboratory
has no alternative water supply, e.g. rain water in storage tanks.
 When intermittent water supplies interrupt cleaning of the laboratory,
decontamination of infectious material, and washing of laboratory ware.
Unreliable water supply-Prevention
 If there is a shortage of piped water, provision is made to get water
supply from other sources or through the water storage tanks.
 The admin department has to be contacted for the same.

Reporting an accident or laboratory related illness

All accidents and laboratory associated illnesses must be reported
immediately and recorded in a Laboratory Accident/Incident form. The
following information is noted:
 Date and time of accident
 Person or persons involved
 Injuries sustained
 Emergency First Aid given and by whom
 Details of follow up actions
 Possible reasons for the accident and preventive action taken.

2. Occupational Injuries

Work related injuries can be categorized as an injury being sustained by:

 Scientific/ technical staff during the collection, handling, processing or
disposal of clinical specimens.
 Housekeeping staff during physical cleaning of the laboratory premises or
during handling of bio medical waste.
 Engineering services personnel during repair/maintenance processes
involving parts of instruments coming in contact with or exposed to clinical
specimens.
 Distribution services personnel during the transport of such specimens to
or from the laboratory.
 Immediate treatment for work related Injuries
 Appropriate first aid measures shall be administrated to the injured.
 The offending needle or broken glass piece should be removed, followed
by ample flushing of the injured part under running tap water. It shall be
insured that the running water has access to the exact injured site.
 In the case of accidental splashing of mucosa of eyes, nose or mouth, the
affected part should be washed with ample volumes of water. The
installed eye wash basin should be used if conveniently accessible or any of
the toilets on the premises.
 If the injury is of the “chemical” rather than a biological sample occurring due
to contact with or exposure to acids, alkalis, alcohols, aldehydes, and etc.,
appropriate first aid measures neutralizing the chemical agent should be
instituted.
 For all injuries that need immediate hospitalization, appropriate arrangements
shall be made by the Safety Officer and Human Resources Manager to
take the patient to the emergency services (Casualty) of the nearest hospital.
 A record of the nearest hospital is available in the list of emergency numbers.

Documentation of Occupational Injury

 All injuries falling into any of the categories enlisted under “Background” shall
be documented in the “work related injuries” register and a written report
shall be submitted to the safety officer within 24- 48 hours of the injury, through
the section head, with the following points clearly mentioned.
 Date and time of occurrence
 Exact nature of the injury
 Extent of the injury/ duration of exposure
 First aid measures administered
 Reason for injury
 Preventive action taken
 The safety officer shall give clear directives regarding the further medical
management and shall ensure that the following measures are complied with
and recorded.
 Follow up steps taken, including the medical treatment, laboratory and
radiological testing etc.
 The staff will be followed up at 6 months for any disability, deformity or
residual effect

3. Emergency preparedness plans

Policy:
The laboratory has an Emergency Preparedness Program designed to
manage the consequences of natural disasters or other emergencies that
disrupt the laboratory’s ability to provide care.

Purpose:
To conduct business normally, it is important for the laboratory to have a
strategy on preparation for emergencies. This plan provides a laboratory
or organizational structure so that the laboratory can effectively prepare
for both external and internal disasters that can negatively affect its
environment of care.

Structure:
 The laboratory plays an important role as a provider of care to the residents
around its locality. The laboratory is ready to assist as needed in case of
community emergency, and as appropriate integrates its Emergency
Preparedness Plan with community disaster plans, as appropriate, to
support the community’s response to a disaster. The laboratory will train its
personnel in this plan.
 The Laboratory Manager, in collaboration with the Safety Officer, will tailor
the laboratory-specific Emergency Preparedness Plan.
 Mitigation activities are those a health care organization undertakes in
attempting to lessen the severity and impact a potential disaster or
emergency may have on its operation while preparedness activities are
those an organization undertakes to build capacity and identify
resources that may be utilized should a disaster or emergency occur.

External Disaster:
A civil catastrophe, either manmade or caused by an act of God. An
external disaster may overwhelm normal facilities. This condition can occur
as a result of fires and explosions, storms, civil disorders, multiple injury
accidents, military action, among other causes

Internal Disaster:
 An event such as a fire or explosion resulting in internal casualties or
circumstances. If the situation requires the evacuation of staff and patient,
such evacuation will be coordinated with emergency service personnel
from the fire and police agencies.
 It is the responsibility of the Laboratory Director to activate the Emergency
Preparedness Plan.
 In the event that total evacuation of the laboratory is necessary, the Safety
officer or his/her designee will assume the responsibility for laboratory
evacuation.
 If an internal disaster disables the laboratory’s essential utility services, the
Laboratory director will determine whether a contracted service will be
used so that reserve utility provisions such as emergency power can be
provided. Emergency power will be limited to providing temporary lighting
so staff can perform essential functions, such as processing certain stat
tests.
 Management of patients in disaster situations:
 If a disaster or an emergency involves the laboratory or staff members, all
less-than-essential services will be temporarily modified or discontinued
until the situation allows for resumption of full program ability.
 The Laboratory director will determine whether these less thanessential
services are to be effected and, if so, when.
 All staff members will be familiar with the overall laboratory Emergency
Preparedness Plan.

4. Evacuation Plan
Planned evacuation will be initiated by the Laboratory director or Safety
officer only.

Evacuation Areas:
The Main Gate on ground floor will be the designated evacuation area and
in case that area is also affected, then the Laboratory director or Safety
Officer will indicate a secondary evacuation area.

Floor Evacuation plan is displayed in all departments.

5. Blood borne pathogens

 The laboratory employees are at an increased risk to potential skin, eye,
mucous membrane, or parenteral contact with blood or other potentially
infectious material and are classified as having occupational exposure to
Blood borne Pathogens
 The collection, accessioning, processing, separating, aliquoting and testing of
the following body fluids: blood and blood components (including control
fluids), semen, vaginal secretions, cerebrospinal fluid, synovial fluid, pleural
fluid, amniotic fluid, unfixed human tissue or organ (other than intact
skin), urine and any body fluid that is visibly contaminated with blood.
 The following job classifications are assigned some or all of the above
duties and therefore subject employees to the risk of occupational
exposure to Blood borne Pathogens: accessioning staff, phlebotomists,
pathologists and pathology assistants, technician performing tests on body
fluids.

Universal safety Precautions: (U.S.P.)

All body fluids must be handled in accordance with the Universal Safety
Precautions that are intended to prevent parenteral, mucous membrane
and non-intact skin exposure of healthcare workers to blood borne
pathogens. Universal safety precautions imply that all body fluids are treated
as if known to be infectious for Human Immunodeficiency Virus (HIV),
Hepatitis B Virus (HBV), or other blood borne pathogens.

a) PPE PROVISION AND USAGE

 All personal protective equipment (i.e. gloves, lab coats, face shields etc.)
shall be removed immediately upon leaving the work area or as soon as if
overtly contaminated (or a tear in case of gloves) and placed in an
 Safety Manual

appropriately designated area or container for storage, washing,

decontamination or disposal.
 Each laboratory personnel should use the following protective equipment (as
required):
 Disposable nitrile gloves
 Laboratory coats
 Closed toe shoes
 Protective face shields / masks wherever applicable
 This protective equipment is available in appropriate sizes to fit all employees
and is either issued to the employee or, as in the case of gloves, is readily
accessible at the work-site.
 Gloves shall be worn when the employee has the potential for the hands
to have direct skin contact with blood or other potentially infectious
materials. In addition, gloves shall be worn when handling any equipment
with the biohazard symbol.
 Disposable (single use) gloves shall be replaced as soon as possible when
visibly soiled, torn, and punctured or when their ability to function as a barrier
is compromised.
 Gloves shall neither be reused nor decontaminated for future use.
 Laboratory workers must be diligent in avoiding contamination of a clean
area by contact with contaminated gloves.
 Gloves should be removed before handling telephones (labelled as “do not
use with gloves”), clean areas or doorknobs etc.
 Laboratory coats shall be worn if there is a potential for soiling of clothes with
blood or other potentially infectious materials. It is compulsory to wear
laboratory coats at all times in the laboratory. Responsibility of the
washing lab coats lies with the admin department of XXXXX. Lab coats will
not be taken home for washing for infection prevention and control
purposes. Washing is done free of cost regularly to ensure clean lab coats.
 Face shields or eye goggles shall be used whenever splashes, spray, splatter,
droplets, or aerosols of blood or other potentially infectious materials may be
generated and there is a potential for eye, nose, or mouth contamination.(i.e.
during washing of glassware, during Elisa procedure). It is required that a
face shield be worn in these areas and while performing any duties where
aerosols may be formed or splattering may occur.
 All employees, who accession, process, test, or directly handle laboratory
specimens shall wear lab coats and gloves.
 All computers, computer key board, computer mouse, door, doors knobs ,
door handle are labelled with “With gloves ” and “without gloves” label

b) Hand Hygiene
The 5 Moments for Hand Hygiene approach defines the key moments when
health-care workers should perform hand hygiene.
This evidence-based, field-tested, user-centred approach is designed to be
easy to learn, logical and applicable in a wide range of settings.
This approach recommends health-care workers to clean their hands
 before touching a patient,

Page 16 of 74
 Safety Manual

 before clean/aseptic procedures,

 after body fluid exposure/risk,
 after touching a patient, and
 After touching patient surroundings.

Hand hygiene is therefore the most important measure to avoid the transmission
of harmful germs and prevent health care-associated infections.
Safety Manual
 Safety Manual

c) Manual Manipulation of needles

 Used needles, blood containing capillary tubes, blood -containing broken
collection tubes and other sharps shall be disposed of in puncture resistant
sharps containers. These containers for ‘SHARPS’ are labelled for the same
purpose with a bio hazard logo .

d) Eating /Mouth Pipetting

 Smoking, vaping, eating, gum chewing, drinking, application of cosmetics
& lip balm, manipulation of contact lenses, and mouth pipetting are
prohibited in all technical work areas.
 Food and drink shall not be stored in refrigerators, freezers, shelves, cabinets
or on countertops or bench tops where blood or potentially infectious
materials are stored.
 Mouth pipetting is prohibited.

e) Specimen Transport Procedure

 All samples from each collection center are transported by visit and
logistics boys in a specially designed transport bag which is strong and is
checked for wear and tear at regular intervals.
 All these bags have a biohazard logo printed on it.
 From collection room samples are transported to accession area by
designated trained staff using bags with bio hazard logo .
 The samples are then accepted by the technicians sitting in this area and
then sorted into different designated trays.
 The respective dept. is called and informed about the arrival of samples in
their department.
 The technicians from each department will take their own department
samples in a transportation labeled plastic trays wearing gloves.

f) Spill Handling
 Spill of either blood or body fluid has to be handled carefully
 If the spill is of significant size, isolate the area and immediately contact
respective senior technician or HOD.
 Use a designated spill kit available with the department.
 The spill kit has the following contents: double pair of gloves,
absorbent material (either cotton roll or tissue roll , container h 10ml 4%
hypochlorite, container with 30 ml tap water/ distilled water, dust pan and
broom, yellow bag, red bag and sharp container with Biohazard logo.
 Put on appropriate personal protective equipment (i.e., double pair of
gloves) and immediately cover the spill with tissue paper/absorbent material
and then pour disinfectant solution (1% hypochlorite).
 Prepare 1% hypochlorite solution, by pouring the 30ml water in 4%
hypochlorite solution container.
 Pour the freshly prepared 1% hypochlorite solution on the spilled surface
in such a manner that the solution flows inward from outside.
 Safety Manual

 Allow 1% hypochlorite solution to remain in contact with the infected surface

for 30 minutes.
 At the end of 30 minutes carefully remove the soaked tissue (wearing gloves)
with the help of dustpan and broom. Always use the broom (provided in
the spill kit) and scoop the material into a plastic dustpan.
 Dispose the collected infectious material in yellow bag.
 Remove gloves and dispose in red bag.
 If broken glass is present, do not try to remove materials (including
broken glass) with bare hands. Always use a small broom (provided in the
spill kit) and scoop the material into it using a plastic dustpan to collect the
material. The collected material should then be disposed into the sharps
container.
 After use the dustpan and broom should be soaked in 1% sodium
hypochlorite solution for at least 2 hours.
 The housekeeping will then be called to clean the area by mopping with
phenyl and water.
 Wash your hands thoroughly with soap and water.
 A training of blood and body fluid spill management is conducted once
annually.

g) Hepatitis B Vaccinations
 All the technical and non-technical staff handling infectious material is
eligible for hepatitis B vaccination.
 The primary course of hepatitis B vaccine is given at 0 day, 30 days and
180 days interval.
 The vaccine is given as an intra muscular injection.
 A series of three intramuscular injections are required to achieve
optimal protection.
 If the vaccination series is interrupted after the first dose, the second
dose should be administered as soon as possible. The second and third
doses should be separated by an interval of at least 2 months. If only the
third dose is delayed, it should be administered when convenient.
 Hepatitis B vaccination is given in the first week of joining the job
 Those staff members who refuse to take the vaccine have to fill
Vaccination refusal form and submit to the quality manager.
 After finishing the third dose of vaccine at 6 months, anti-HBV titer is done at
a minimum of 40 days after completion of the third dose.
 A titer of more than 10 I.U/ml is considered adequate protection. A record of
vaccination of the staff (FF Hepatitis B vaccine record and hepatitis B vaccine
summary record)) is with the laboratory manager.
 In case the titre is less than 10 I.U/ml, a repeat vaccination of the staff needs
to be done and anti-HBV titers repeated. If the titers do not increase, the
employee is labeled as non- responder and is then recruited into work that
does not involve handling of patient samples.

Page 20 of 74
 Safety Manual

 The immunization record should have Vaccination dates with employee

signatures and also Anti HBsAg titer report.

h) H1N1 Vaccination
 H1N1 collection will be done by designated trained staff at each Centre.
 A record of the training for collection needs to be documented.
 The designated staff will be vaccinated annually once for H1N1. The
vaccine will be distributed from Purchase department to respective center
managers after the approval of Training & Development Team.
 Vaccine Name: Inactivated Influenza Vaccine (Surface antigen IP)
 Vaccine Storage: At 2-8°C
 Vaccine administration: Intra muscular
 Person responsible for vaccination: Physician at each center
 Vaccine dosage: 0.5 ml prefilled syringe for single use
 Record of vaccination needs to be maintained by center manager in FF-
H1N1 Vaccine Summary Record

i) COVID-19 Vaccination
 Staffs are encouraged for the vaccination of both the dosages.
 Vaccinated staff data is maintained with the respective managers/HODs in
FF-COVID Vaccine Summary Record (as per MoHFW)

j) Tetanus Vaccination
 Basic biomedical safety training is given to all the staff so that he/she must be
aware of lab waste and working materials. They should be protected against
Tetanus.(ICMR, GCLP, 2021,4(h)- Bio Medical Waste Management Rules
- 2016)
 All the staff involved in handling of Bio Medical Waste will be vaccinated
once for Tetanus on joining.
 Respective teams will procure the vaccines through Purchase inventory
management system as per the requirement after the approval of HOD.
 Vaccine Name: BETT Tetanus Toxoid vaccine
 Vaccine Storage: At 2-8°C
 Vaccine administration: Intra muscular
 Person responsible for vaccination: Physician at each center
 Vaccine dosage: 0.5 ml prefilled syringe for single use
 Record of vaccination needs to be maintained in FF-Tetanus Vaccine
Summary Record.

Page 21 of 74
 Safety Manual

k) Viral Exposure (Post exposure prophylaxis plan -PEP)

XXXXX has established a post exposure prophylaxis plan –PEP: evaluation
and treatment program that includes testing to determine whether
infection with HIV, HBV and/or HCV has occurred, and includes follow-up
treatment and counselling.
Policy is that any employee having an occupational (workplace) exposure
incident shall be offered the post-exposure program. This plan is prepared as
per the NACO Guidelines.

POST EXPOSURE PROPHYLAXIS

PLAN PROCEDURE

a. For splash to the eyes, nasal mucosa or mouth

Wash the exposure site with large amounts of tap water using water available at an
eyewash station or any sink. Flush the eye, nasal mucosa or mouth thoroughly with
water.

b. Needle stick injury

If an employee has a percutaneous injury (needle stick or cut), the senior

technician/lab manager shall be informed IMMEDIATELY to take the
following recommended actions:

 Wash thoroughly with water.

 The employee shall immediately notify senior technician/lab manager of the
exposure.
 The staff is given Duovir tablet (available at every center) within 2 hours of
exposure, after consultation and approval of Safety Officer (if bleeding happens
without pressing the site).
 The same should be escalated to Safety Officer, immediately. Additionally,
the senior technician/lab manager should notify the Safety officer regarding
the accident via email.
 Verify the employee has been directed to seek medical attention.
 When recommended, Post exposure prophylaxis(PEP) should be started as soon
as possible, preferably within 2 hours. Initiating treatment after 72 hours
of exposure is not of much use.
 The decision to start PEP is made on the basis of determination of extent of
exposure and HIV/HBsAg/HCV status (of the source).

 Factors that influence Risk of Infection

 Various epidemiological and laboratory studies have shown that the risk of
infection, following exposure, varies with the type of exposure:
 Type of needle (hollow bore vs. solid)
 Device visibly contaminated with patient’s blood
 Depth of injury

Page 22 of 74
 Safety Manual

 The amount of blood involved in the exposure

 The amount of virus (viral load) in the exposed blood/body fluid at the
time of exposure.
 Timely (<2 hours and up to 72 hours) availability and efficacy of the PEP.
 Testing: Both the employee and source specimen should be tested for
HIV antibody, HBS antigen and HCV antibody.
 Assessing the HIV status of the source of exposure
HIV status of the source Definition of risk in source

HIV negative Source is not HIV infected(but consider HBV and HCV)

Low risk HIV positive and clinically asymptomatic

High risk HIV positive and clinically symptomatic

Status of the patient is unknown and neither the patient

Unknown nor his/her blood is available for testing

ASSESSING THE NATURE OF EXPOSURE

Category Definition and example

Mild Exposure Mucus membrane/non-intact skin with small volume, e.g., a

superficial wound (erosion of the epidermis) with a plain or low

calibre needle; contact with the eyes or mucous membranes;

Subcutaneous injections following small bore needles.

Moderate Mucus membrane/non-intact skin with large volumes or

Exposure

percutaneous superficial exposure with solid needle (e.g., a cut

or needle stick injury penetrating gloves).

Severe Exposure Percutaneous with large volume, e.g., an accident with wide bore
needle (>18G) visibly contaminated with blood

; a deep wound

(haemorrhagic wound and/or very painful); transmission of a

significant volume of blood; an accidental injury with material,

which has previously been used intravenously or intra-arterially

 The staff is given antiretroviral medication (available at every centre) within

2 hours of exposure if recommended as per the above table.
 The staff is then referred to a medical specialist – M.D. Medicine.

Version No/Date: XX Page 23 of 74

 Safety Manual

 The treatment needs to be taken as per the treating physician’s instructions.

 The cost of treatment and testing will be borne by the respective centre.
 An email should be sent to the Safety officer regarding the incident, registration
numbers of the staff and the source and the reason for needle prick.
 A record of the incident is maintained in the needle stick injury file by the safety
officer.
 The cost of treatment for the staff will be borne by each individual centre.
 The concerned staff can buy the medicines and give the receipt to the centre.
The medicines have to be taken for 1 month.
Also the patient's and staff's blood will be collected for HIV, HbsAg, and HCV as
baseline tests. A follow up will be taken for HIV/HCV & HbsAg post 6 months.

Determination of HIV status code (HIV SC)

1. HIV Status of exposure source

HIV HIV STATUS SOURCE
UNKNOWN UNKNOWNN

HIGH titre exposure

HIV Low titre Exposure e.g. Advanced AIDS,
(e.g. A post-
NEGATIVE primary HIV infection /
exposure program.
high viral load or low
Symptomatic/
high CD4 count) CD4 count
HIV SC
HIV SC 1 HIV SC 2 UNKNOWN

2. Exposure Code (EC)

IS THE SOURCE MATERIAL BLOOD, BODY FLUID, OTHER
POTENTIALLY INFECTIOUS MATERIAL (OPIM) OR AN
INSTRUMENT CONTAMINATED?

NO PEP
NO YES

OPIM, BLOOD/ BODY FLUIDS

TYPE OF EXPOSURE

INTACT PER CUTANEOUS EXPOSURE

SKIN
Version No/Date: 3.4/13-05-2022 Page 24 of 74
 Safety Manual

MUCUS MEMBRANE /SKIN OR

INTEGRITY COMPROMISED?

LESS SEVERE(E.g. Solid MORE SEVERE (e.g.

needle, superficial Large bore hollow
LARGE VOLUME(E.g. needle, Deep
scratch)
several drops, major Puncture, visible blood
SMALL VOLUME (e.g.
splash/ longer duration
few drops, /short several minutes or on device or needle
duration more used in patient’s artery/
vein

EC 1
EC 2 EC 3 EC 3

 Determination of PEP Recommendation

EC HIV SC PEP RECOMMENDATION Duration

1 1 Not warranted

1 2

2 1
Recommended
2 2 28 days

3 1 or 2

Consider PEP, if HIV prevalence is high in the

2/ 3 Unknown
given population & risk categorisation

Page 25 of 74
 Safety Manual

Management in case of exposure with HCV :

Page 26 of 74
 Safety Manual

Management in case of exposure with HBsAg:

Postexposure Management
Initial Postexposure Management
Procedure is followed for testing known source, including obtaining informed
consent, in accordance with applicable laws. When a source is unknown (e.g., as
occurs from a puncture with a needle in the trash), the exposed HCP is managed as
if the source patient were HBsAg-positive.
Laboratory should ensure that HCP have timely access to postexposure
management and prophylaxis, including HBIG and HepB vaccine. For exposed
HCP thought to be susceptible to HBV infection, HBIG and HepB vaccine
should be administered as soon as possible after an exposure when indicated.
The effectiveness of HBIG when administered >7 days after percutaneous,
mucosal, or non-intact skin exposures is unknown. HBIG and HepB vaccine can be
administered simultaneously at separate injection sites.
Anti-HBs testing of HCP who received HBIG should be performed after anti-HBs
from HBIG is no longer detectable (6 months after administration). Anti-HBs testing
should be performed using a method that allows detection of the protective
concentration of anti-HBs (≥10 mIU/mL)

Postexposure management of health-care personnel after occupational percutaneous and mucosal

exposure to blood and body fluids, by health-care personnel HepB vaccination and response status

Postexposure
Postexposure testing Postvaccinati
Health-care personnel prophylaxis
on serologic
status Source patient HCP testing
HBIG* Vaccination testing†
(HBsAg) (anti-HBs)
Documented responder§
after complete series (≥3 No action needed
doses)
HBIG x2
Documented Positive/unkno separate
—** — No
nonresponder wn d by 1
after 6 doses month
Negative No action needed
Positive/unkno <10mIU/mL
HBIG x1 Initiate
wn **
Response unknown after Yes
revaccinati
3 doses Negative <10mIU/mL None
on
Any result ≥10mIU/mL No action needed
Positive/unkno Complete
Unvaccinated/incomplet —** HBIG x1 Yes
wn vaccination
ely vaccinated or
vaccine refusers Complete
Negative — None Yes
vaccination

Page 27 of 74
 Safety Manual

Abbreviations: HCP = health-care personnel; HBsAg = hepatitis B surface antigen;

anti-HBs = antibody to hepatitis B surface antigen; HBIG = hepatitis B
immune globulin.
* HBIG should be administered intramuscularly as soon as possible after exposure
when indicated. The effectiveness of HBIG when administered >7 days after
percutaneous, mucosal, or nonintact skin exposures is unknown. HBIG dosage is
0.06 mL/kg.
Should be performed 1–2 months after the last dose of the HepB vaccine series (and
4–6 months after administration of HBIG to avoid detection of
passively administered anti-HBs) using a quantitative method that allows
detection of the protective concentration of anti-HBs (≥10 mIU/mL).
A responder is defined as a person with anti-HBs ≥10 mIU/mL after ≥3 doses of
HepB vaccine.
A nonresponder is defined as a person with anti-HBs <10 mIU/mL after ≥6 doses of
HepB vaccine.
** HCP who have anti-HBs <10mIU/mL, or who are unvaccinated or
incompletely vaccinated, and sustain an exposure to a source patient who is
HBsAg-positive or has unknown HBsAg status, should undergo baseline testing
for HBV infection as soon as possible after exposure, and follow-up testing
approximately 6 months later. Initial baseline tests consist of total anti-HBc;
testing at approximately 6 months consists of HBsAg and total anti-HBc.

Managing Vaccinated HCP

For vaccinated HCP (who have written documentation of a complete, ≥3-dose

HepB vaccine series) with subsequent documented anti-HBs ≥10 mIU/mL, testing
the source patient for HBsAg is unnecessary. No postexposure management for HBV
is necessary, regardless of the source patient's HBsAg status.
For vaccinated HCP (who have written documentation of HepB vaccination) with
anti-HBs <10 mIU/mL after two complete, ≥3-dose HepB vaccine series, the source
patient should be tested for HBsAg as soon as possible after the exposure. If the
source patient is HBsAg-positive or has unknown HBsAg status, the HCP should
receive 2 doses of HBIG . The first dose should be administered as soon as possible
after the exposure, and the second dose should be administered 1 month later.
If the source patient is HBsAg-negative, neither HBIG nor HepB vaccine is
necessary.
 If the HCP has anti-HBs <10 mIU/mL and the source patient is HBsAg-positive or
has unknown HBsAg status, the HCP should receive 1 dose of HBIG and be
revaccinated as soon as possible after the exposure. The HCP should then
receive the second 2 doses to complete the second HepB vaccine series (6
doses total when accounting for the original 3-dose series) according to the
vaccination schedule. To document the HCP's vaccine response status for
future exposures, anti-HBs testing should be performed 1–2 months after the last
dose of vaccine.

Page 28 of 74
 Safety Manual

 If the HCP has anti-HBs <10 mIU/mL and the source patient is HBsAg-
negative, the HCP should receive an additional HepB vaccine dose, followed
by repeat anti-HBs testing 1–2 months later. HCP whose anti-HBs remains
<10 mIU/mL should undergo revaccination with 2 more doses (6 doses
total when accounting for the original 3-dose series). To document the
HCP's vaccine response status for future exposures, anti-HBs testing should be
performed 1–2 months after the last dose of vaccine.
 If the HCP has anti-HBs ≥10 mIU/mL at the time of the exposure, no
postexposure HBV management is necessary, regardless of the source patient's
HBsAg status.

Managing HCP Who Lack Documentation of Vaccination, are Unvaccinated or

Incompletely Vaccinated

For unvaccinated or incompletely vaccinated HCP (including those who refused

vaccination), the source patient should be tested for HBsAg as soon as possible
after the exposure. Testing unvaccinated or incompletely vaccinated HCP for
anti- HBs is not necessary and is potentially misleading, because anti-HBs ≥10 mIU/mL
as a correlate of vaccine-induced protection has only been determined for
persons who have completed an approved vaccination series
 If the source patient is HBsAg-positive or has unknown HBsAg status, the HCP
should receive 1 dose of HBIG and 1 dose of HepB vaccine administered as
soon as possible after the exposure. The HCP should complete the HepB
vaccine series according to the vaccination schedule. To document the
HCP's vaccine response status for future exposures, anti-HBs testing should
be performed approximately 1–2 months after the last dose of vaccine.
Because anti-HBs testing of HCP who received HBIG should be performed
after anti-HBs from HBIG is no longer detectable (6 months after
administration), it will likely be necessary to defer anti-HBs testing for a
period longer than 1–2 months after the last
vaccine dose.
— HCP with anti-HBs ≥10 mIU/mL after receipt of the primary vaccine series
are considered immune. Immunocompetent persons have long-term
protection and do not need further periodic testing to assess anti-HBs
levels.
— HCP with anti-HBs <10 mIU/mL after receipt of the primary series should be
revaccinated. For these HCP, administration of a second complete 3-dose
series on an appropriate schedule, followed by anti-HBs testing 1–2 months
after the third dose, usually is more practical than conducting serologic testing
after each additional dose of vaccine. To document the HCP's vaccine
response status for future exposures, anti-HBs testing should be performed 1–
2 months after the last dose of vaccine.
 If the source patient is HBsAg-negative, the HCP should complete the
HepB vaccine series according to the vaccination schedule. To document
the HCP's vaccine response status for future exposures, anti-HBs testing
should be performed approximately 1–2 months after the last dose of
vaccine.
Page 29 of 74
 Safety Manual
— HCP with anti-HBs ≥10 mIU/mL after receipt of the primary vaccine series are

Page 210 of
 Safety Manual

considered immune. Immunocompetent persons have long-term

protection and do not need further periodic testing to assess anti-HBs levels.

Testing of HCP Exposed to an HBsAg-Positive or Unknown Source

HCP who have anti-HBs <10 mIU/mL, or who are unvaccinated or incompletely
vaccinated, and who sustain a percutaneous, mucosal, or nonintact skin
exposure to a source patient who is HBsAg-positive or has unknown HBsAg
status should undergo baseline testing for HBV infection as soon as possible
after the exposure, and follow-up testing approximately 6 months later. Testing
immediately after the exposure should consist of total anti-HBc, and follow-up
testing approximately 6 months later should consist of HBsAg and total anti-HBc.
Vaccine Nonresponders
Vaccinated HCP who’s anti-HBs remains <10 mIU/mL after revaccination (i.e., after
receiving a total of 6 doses) should be tested for HBsAg and anti-HBc to
determine infection status. Those determined not to be HBV infected (vaccine
nonresponders) should be considered susceptible to HBV infection. No specific work
restrictions are recommended for vaccine nonresponders.

The test needs to be registered at the respective centre and the patient ID
number of the patient and staff should be emailed to the Safety officer-(Name) at
(email id) along with the reason for needle prick. The cost of consultation with
the doctor will also be borne by each individual center.
 Safety Manual

l) TB Exposure
Tuberculosis Exposure control plan

Individuals who have patient contact or handle body fluid specimens that
may contain mycobacteria should undergo tuberculosis screening at the
time of their pre-placement examination and periodically as required by
this plan.

 Pre-placement Screening: The PPD conversion status of the employee

must be noted at the time of pre-placement examination. A chest X-ray
shall be obtained and a symptom review shall be performed. If symptoms or
chest X- ray are suggestive of tuberculosis, a referral will be made for
evaluation and treatment. The individual will be placed on medical hold
until deemed non- infectious. For employees with a history of tuberculosis,
information shall be obtained regarding the age at diagnosis, duration of
the treatment and medication.

 Periodic TB Screening: Training and information shall be provided to

ensure employee knowledge of the hazard of TB transmission, its signs and
symptoms, medical surveillance and compliance with this plan. Periodic
screening shall be performed every six months for employees in high-risk
departments like mycobacteriology and annually for other “at risk”
employees. This screening will be done on a departmental basis. This would
be done in the following manner:

 The following tests will be performed every 6 monthly and if the at risk
persons exhibit symptoms of TB in the form of cough for more than 2
weeks not responding to antibiotics, low grade fever, weight loss and loss
of appetite. The following test will be done :
 CBC, ESR
 XRAY CHEST
 Mantoux test
 Physical examination

 Annual chest X-ray and symptom review: If the chest X-ray is positive for
suspected active TB, or if the employee has a negative chest X-ray with
symptoms of TB, the employee shall be immediately withdrawn from his work
area. If the employee is found to have active TB, the employee shall
remain off work until documentation from the employee’s treating health
care provider is received stating that the employee is non-infectious.

 Symptomatic Individuals: Seniors employed with affected department should

be suspicious of individuals exhibiting symptoms of infectious or active TB.
Symptoms of pulmonary TB include night sweats, weight loss, chronic
coughing, blood in expectoration, fatigue and chest pain. Suspect
infectious
 Safety Manual

individuals shall be referred to the Safety Officer for out of turn

Tuberculosis screening

 Immuno-compromised employees: Severely immune-

compromised employees will be advised to avoid exposure to TB.
Employees should be advised of options for the severely immune-
compromised, to voluntarily transfer to areas and activities in which there is
a reduced risk of exposure to TB. This should be a personal decision for the
employee after being informed of the risk to her/him and evaluating her/his
job commitment and satisfaction. Confidentiality will be maintained.

m)Sterilizing Device Monitoring:

 A record of chemical indicator strip placed in each run of both autoclaves:
Discard and media preparation is to be maintained.
 Chemical indicator strips reflecting sporicidal conditions are used. A color
change from olive green to dark Brown/ Black is to be observed and
recorded.
 A record of Biological indicators (Bacillus stearothermophillus spore indicator
strip) is placed weekly in the discard autoclave and the records are
maintained. The spore indicator vial is placed at 56 degree C for 2 days
and the color change is observed and recorded.

6. Fire Prevention and Protection

Classes of Fire (As Per Indian Standard)

 Class A-fires involving solid materials such as wood, paper or textiles.

 Class B-fires involving flammable liquids such as petrol, diesel or oils.

 Class C-fires involving gases & live electrical apparatus.

 Class D-fires involving metals.

 Class K-fires involving cooking oils

What do you do in case of fire

 Safety Manual

How to use a Fire Extinguisher

a) Fire Separation
The laboratory is properly segregated from inpatient areas.

b) Fire Exit:
 Laboratory is having two permanent exit routes to permit prompt evacuation
of all staff during emergency
 Both the exit routes are away from each other
 The fire exit route is free of inflammable/combustible material, furnishing
& decoration.
 The fire exit route is not obstructed by materials, equipment’s & locked doors.

c) Annual Fire Drill:

 Fire drill will be conducted once a year. The drill will be in practical and not on
paper only. The fire drill will comprise of the following events.
 The moment the alarm goes off, the safety officer or any designated staff will
inform the nearest fire station about the event.
 All emergency numbers including fire station contact number is displayed
in the lab.
 All personnel will stop whatever work they are doing and start walking,
creating human chain (not running) towards the emergency exit door in a
single or double file without jostling. The staff will walk through the stairs and
not use the lift.
 The staff will collect on the ground floor near exit gate.
 A count of the staff will be taken then.
 The record of fire drill conducted by the authorized personnel is available with
the quality department/ safety officer.

d) Fire detection/Alarm:
 A fire alarm system has been installed in the lab which will get activated in
case there is fire due to any reason and smoke is getting generated.
 A certificate of compliance, working and training of all staff on the system
is documented annually.
 Safety Manual

e) Fire Extinguishers:
 Apart from the above fire fighting system, the lab is also equipped with
fire extinguishers of ABC-rated and CO2 type.
Fire extinguisher Class of fire it
Usage
type extinguishes
Generally used in
Dry Chemical powder A, B & C
laboratory
To be used for electrical
CO2 B&C
fire risks

 These fire extinguishers have been placed at strategic locations in the lab
area to face any eventualities. The certificate of compliance of these
extinguishers is available with the Admin department. A record of training and
use of fire extinguisher of each staff is available with the Admin department/
safety officer.
 Fire extinguishers should be placed in lab such that distance for staff to
any extinguisher is 75 feet (22.9 m) or less(As per OSHA 1910.157(d)(2)
guidelines)
 CO2 type of fire extinguisher should be placed near to the laboratory
equipments.
Nearest Fire Station : Address

All the Emergency lists should be displayed at the centres and Labs.

7. Electric Safety
 All personnel must know the location of master switches and circuit
breaker boards. Do not attempt to repair any instrument while it is still
plugged in.
 Plugs or cords that are broken, frayed, or worn should not be used.
 Outlets must not be overloaded. Never use gang-type plugs.
 All cord and plug-type electrical equipment should have grounded power
cords and plugs. All shocks, including small tingles, must be immediately
investigated.
 Extension cords should be used only in compliance with the overall lab
policies and procedures.

8. Chemical Safety

a) Chemical Hygiene plan (available in detail in sop on chemical hygiene

plan) In the event that any employee is exposed to a hazardous chemical,
following procedure must be followed.
 Decontamination should be conducted if there was contact with the
clothing or skin. This may include removing any contaminated clothing
and rinsing/washing the contaminated area with generous amounts of water.
 In an emergency situation, seek medical attention immediately,
 Safety Manual

 If the exposure does not constitute an emergency, the employee must be

told that they are entitled to a medical evaluation at no cost to them.

b) MSDS
 All reagents on the chemical inventory have an MSDS.
 MSDS must be available in each laboratory section. The MSDS shall be
arranged in alphabetical order by chemical along with a chemical
inventory of the laboratory section. The laboratory shall rely on the
chemical manufacturer’s information to ascertain whether or not the
chemical is hazardous.
 The MSDS for all chemicals is available online in the SOP folder at each
desktop in the lab

c) Chemical Precautionary Labels

 Original manufacturer’s containers have appropriate labelling.
 All secondary containers of hazardous chemicals must be labelled with a
minimum of the identity of the reagent as its name appears on the bottle.
Furthermore, all secondary container labelling shall include any unique
warnings such as “ACID”, CARCINOGEN”, etc.
 Labels shall not be removed or defaced.

 All acids are labelled with a corrosive logo

d) Flammable, Acid Base & Volatile solvent Storage

 Storage on bench tops and in hoods may cause potential exposure to fire
and spills and must be avoided
 The entire chemical is stored in separate cabinet which is below the eye level.

 The cabinet is labelled as inflammable reagents with a logo

 The cabinets are well ventilated.
 All reagents are stored on sand.
 Safety Manual

9. Environmental Safety

a) Laboratory Ergonomics Program

XXXXX takes all reasonable precautions to protect the health and safety of its
employees, the public, and the environment. As part of this commitment, XXXXX
has implemented the Ergonomics Program, whose primary objective is to
prevent injuries and illnesses in the workplace. Other components of the
program include work area evaluation, selection and use of appropriate
equipment, and education and training.

This document describes the XXXXX Ergonomics Program and its components.
Contained herein are precautions for preventing upper-extremity injuries and
illnesses, roles and responsibilities for all workers, basic ergonomics principles and
practices, and the resources available to workers and supervisors for identifying and
resolving ergonomic problems.

Failure to use (or improper use of) the precautions outlined in this document
can lead to many different musculoskeletal illnesses and injuries, some of which can
be debilitating. A musculoskeletal illness or injury is defined as an illness or injury of
the muscles, tendons, ligaments, joints, cartilage, peripheral nerves, vascular
system, or other related soft tissue.

The term "ergonomics" refers to the relationship between individuals and their work
environment. The problems addressed by ergonomics include improper "fit" of
the workplace, poorly designed or improper tools, and poor body mechanics
when lifting or performing repetitive tasks (including computer keyboard use).

BACKGROUND

(A) Identifying a Possible Ergonomic Problem

Many ergonomic disorders are felt as strains and sprains. Acute or chronic
muscle strain can be an indication that the capacity of the body to
accommodate physical stressors has been exceeded. Chronic strain and
cumulative trauma disorder (CTD) result from less-intense stresses that
accumulate over time, reducing the rate of recovery of the musculoskeletal system.

Acute Muscle Strain

The signs and symptoms of acute muscle strain generally may include pain within 24
hours of an injury to the musculoskeletal system.
Most acute muscle strain injuries can be prevented. To prevent injuries:
 Safety Manual

 Use mechanical devices or additional personnel when lifting and moving

heavy loads.

 Use proper body mechanics.

 Establish limits for lifting heavy objects.

 Avoid excessive fatigue from repeated forceful activities.

Workers should report symptoms of acute muscle strain to their work supervisor and
then report to the Health Services Department.

Cumulative Trauma Disorder

The signs and symptoms of CTD of the upper extremities include pain,
numbness, and tingling of the fingers, wrist, elbow, or shoulder. Chronic back
and neck problems may result in pain, numbness, or tingling that radiates to the
arms or legs, as well as limited back motion. Doing the following usually can
prevent these problems:
 Use ergonomically designed tools and workplaces (e.g., furniture that has
adjustment flexibility and allows for proper posture).

 Educate workers to adhere to ergonomically appropriate work habits (e.g.,

maintaining the proper posture and using a light touch when doing keyboard
work).

 Vary physical activities appropriately to allow frequent, short rest periods

during which tendons and muscles are not subjected to repetitive strain or
sustained contraction.
It is extremely important for workers to report any recurrent symptoms of CTD
(e.g., pain, numbness, tingling, or tenderness) to their work supervisor and the
Health Services Department.

(B) Ergonomic Design and Practices

(a) Computer Ergonomics

A frequent contributor to CTD is improper configuration and use of computer
workstations. Changes to a workstation may require only repositioning furniture or
equipment or purchasing ergonomically appropriate replacements. Figure 1 shows
a well-designed computer workstation.
 Safety Manual

Figure 1. A well-designed computer workstation.

 A chair should have an adjustable back that provides support for the
lumbar region of the back and trunk. Armrests should be of a padded
material and adjustable in height. The seat pan should be large enough to
be comfortable.
 A work surface should be large enough to accommodate all computer
equipment, including a wrist rest in front of the keyboard and input
device. Sufficient room should be provided under a work surface to allow
free leg movement. The height of the work surface should allow the
forearms to be parallel with the floor while working at the computer.
 A keyboard and input device (mouse or trackball) should be at the same
level and in front of the operator. The height of the keyboard and input
device should allow the operator to position his/her forearms and hands
parallel to the floor during operation.
 A terminal (i.e., monitor) should be located directly in front of the
operator, and the top of the screen should be approximately at eye level
or slightly lower.
 Vision is a critical part of the workstation composition. An annual eye
examination is recommended to ensure that any changes in vision are
detected and corrected.
 Posture:Most of the new crops of keyboard designs are aimed directly at
the posture issue. Improper postures involving prolonged, non-neutral
positions of the joints may stretch, compress, or otherwise stress tendons,
nerves, or other tissues.

Pipettes ergonomics guidelines

The use of manual Pipettes has been associated with a high prevalence of
musculoskeletal disorders of the hand, wrist, forearm, shoulder and neck among
laboratory workers.
More specifically, risk factors for use of traditional pipettes include Repetitive motion
and excessive force of the thumb ,Awkward postures of the hand, wrist, arms
and shoulders.
 Safety Manual

In XXXXX, Ergonomically designed pipettes are used to minimize the problems

of musculoskeletal disorders of the hand.

Seating Ergonomics
1. Back pain is as mystifying today as it was decades ago. Despite excellent
tests and procedures, modern back specialists admit that up to eighty
percent of all cases have no clear physiological causes. In fact, many pain-
free people show bulging or herniated discs in x-rays.
2. There is little agreement on how to do lifting with little risk.
Lifting with the legs is easy on the back, but hard on the legs and muscles. Lifting
with the back puts strain on the disks but is less fatiguing.
3. People who sit for long periods are at risk for back disorders.
The two greatest problems seem to be 1) sitting upright or forward, and 2)
not changing position.
4. An upright posture with a ninety-degree hip position is actually unhealthy, from
the perspective of the intervertebral discs. For a number of reasons, the discs
experience more pressure and the pressure is more lopsided than while standing.
So it's a good idea to sit with the hip joints somewhat straightened.
5. Even if the hip joints aren't somewhat straightened, sitting in a reclined posture
is more healthy than sitting upright. This is because reclined sitting puts
more of your weight onto the chair's backrest.
All sitters should move around. In addition to helping the muscles relax and
recover, this alternately squeezes and unsqueezes the intervertebral discs, which
results in better filtration of fluids into and out of the cores of the discs. Discs
stay plumper and, in the long run, healthier. One implication: chairs should
follow the sitter as he/she changes posture.
6. The most important chair adjustments are
 Seat height from the floor - the feet should be able to rest flat on the
floor. (However, this doesn't mean the feet should always be flat on the floor.
Legs should be free to stay in different positions).
 Depth from the front of the seat to the backrest - sitters should be able to use
the backrest without any pressure behind the knees.
 Lumbar support height - every person is shaped differently.

Hand and arm ergonomics

There are many kinds of CTD (Cumulative Trauma Disorder) medical conditions that
have ergonomic causes among office workers, including carpal tunnel syndrome
and various kinds of tendon inflammation.
UV radiation ergonomics

The laboratory staff may be exposed to UV radiation in the form of UV fluorescence

from the fluorescence microscope. This can be avoided by using the
fluorescence shield provided by the manufacturer.
Another source of UV exposure could be the laminar flows. The scientific staff
and the housekeeping staff should take care not to open the glass hood
whenever the UV light is on. A ‘DO NOT OPEN’ sign should be posted on the laminar
flow.

Management of ergonomic problems .

 Respect pain. If an activity causes pain or discomfort, stop and evaluate

the activity to look for alternative approaches. Change positions if the
activity is causing pain or discomfort.
 Alternate tasks during the workday to interrupt repetitive activities.
 Keep the wrists in the neutral position whenever possible.
 Use two hands whenever possible, even when handling light objects or
doing small tasks.
 Make several trips with lighter loads. Use a cart or dolly, if necessary.
 To avoid the use of a sustained, forceful grip, use a vice, clamp, or jig to
stabilize objects.
 Alternate Work Periods (five minutes for every 30 minutes of work is necessary.
Program Evaluation

The XXXXX Ergonomics Program is evaluated periodically to determine whether

established objectives are being met and whether revisions to the program are
necessary. The program's primary objective is to reduce ergonomic injuries
and illnesses in the workplace. The relevant indicators are monitored and evaluated.

Education and Training

Education and training are key aspects of the XXXXX Ergonomic Program.
Supervisors and workers should receive sufficient information and education to
recognize ergonomic risk factors, to understand the nature of ergonomic injuries
and illnesses, and to be aware of potential corrective measures and the
resources available.

b) Excessive Noise:
The laboratory will protect the working personnel from excessive noise
levels.

c) Emergency Eyewash
 Eye wash station is located in lab so that it takes minimum time to reach from
any department.
 Eye wash station is kept free of obstacles blocking their use.
 Eyewash is capable of delivering 1.5 litres/minutes for 20 minutes.
 Functioning correctly & weekly check is done.
 Provide the quality and quantity of water that is satisfactory for
 emergency washing purposes.
 Signage for eye wash station location is placed which is visible to all staff.
 Safety Manual

 In the event of a chemical splashing into eye(s) or on skin, the affected

area(s) is immediately flushed with running water for at least 20 minutes/ eye
wash facility.
 For labs where eye wash is not available, a wash basin has been provided.

10. Other Hazards

a) UV Light Exposure:
UV light is present inside the Biosafety cabinets of microbiology and molecular
department. All these cabinets will have the warning sign regarding the
hazards of U.V. lights.

b) Latex Allergy:
The laboratory has a policy to protect the personnel from allergic reactions
from exposure to natural rubber latex in gloves hence laboratory has
mandated the rule of using only nitrile gloves. Powdered gloves are no longer
provided to any employees.

c) Dry Ice Hazards & Safe Handling:

 PURPOSE & SCOPE: This procedure describes methods for safely using,
storing, and handling dry ice.
 HAZARD DESCRIPTION:
1. Handle dry ice with appropriate insulated gloves. Using bare hands
can result in burns/frostbite to the skin in a short period of time.
2. Use of dry ice in poorly ventilated areas can result in the depletion of the
oxygen level resulting in asphyxiation.
3. Placing dry ice into a tightly sealed container can produce sufficient
gas build up to cause an explosion.

 RESPONSIBILITIES:
1. Logistics
2. Technician
3. Accession Department

 GENERAL SHIPPING REQUIREMENTS:

1. Dry ice must be packaged in containers that allow the release of CO2
gas.

 PERSONAL PROTECTIVE EQUIPMENT (PPE): Safety goggles, Heavy duty

gloves, lab coat or lab apron must be worn when handling dry ice.

 DISPOSAL OF UNNEEDED DRY ICE:

1. Let the unused portion sublimate (recommended for well‐ventilated
areas because it will happen over several days and ventilation will
take care of the gas liberation).
2. Never dispose of dry ice in a sink, toilet or other drain.

Page 41 of 74
 Safety Manual

3. Never dispose of dry ice in the trash or garbage

4. Never leave surplus dry ice in the corridors.

 REFERENCES: OSHA Quick Fact on Dry Ice

d) First aid box & Spill kit:

 First aid box with thrombophob ointment, bandages, Dettol, Soframycin,
Combiflam etc. should be available.
 Spill Kit should be readily available with Hypochlorite solution & water,
sharp container, gloves, small broom, dust pan, tissue paper & yellow bag.

11. Waste Disposal

All the liquid waste generated from the laboratory are hazardous to heath,
hence it has to be discarded off in appropriate manner as per the local
biomedical waste guidelines.

a) Liquid waste disposal

Instrument generated waste

 Waste to be collected in the empty can which is preloaded with 1/10th
quantity of 1% Sodium hypochlorite.
 Once the container is 3/4th full, remove the container and discard in the
sink designated where ETP is installed for disposal.

P.N: For CRL, liquid generated waste is directly connected to the ETP.
The above procedure is not applicable for CRL; please refer to the
individual ETP SOP.

Page 42 of 74
 Safety Manual

Disposable Micro tips

 All the disposable micro tips have to be discarded in 1% Sodium
hypochlorite small bins and discarded on daily basis.
 Hypochlorite water has to be drained and the tips have to be discarded
in red bag.
 This waste has to be discarded in sink designated where ETP is installed
for disposal.

Effluent Treatment Plant

Purpose:
Effluent Treatment Plant (ETP) is installed at all the processing labs. ETP
ensures that the contaminated water from instruments get treated and become
reusable before being released back to nature.

Process to be followed:
 Instrument generated waste will be discarded in the sink below which the ETP is
installed only.
 Check the Dosing Tank level.
 Wear pair of gloves while handling.
 Prepare the hypo solution. Method of preparation: Prepare 1:80 dilution Take
19.75 liters of water + 250 ml hypochlorite(0.25 liters)-
35% Sodium hypochlorite to be used for dilution.
 Start the dosing pump 5 minutes before dumping of liquid waste.
 Once dumping is over, switch off the dosing pump after 10 minutes.
 Keep the dosing pump knob on 100%.
 Precautions: Do not run the Pump Dry.
 Clean the bag assembly once in every 15 days.

Reference:
Refer to the ETP SOP for other details.

Chemical waste (Xylene, formalin)

 For Histopathology, Only quantities 250 ml at a time can be disposed of in
the sink drain along with the running tap water.
 Continue to run cold water for several minutes after completion.
 Dispose of water-soluble organic solvents (methanol, acetone)
as described above.
 For water-insoluble organic liquids, only quantities less than 100 ml can be
disposed of as described above.

b) Biohazard Disposal Containers

 Biohazard bags are place in closed lid dustbin to prevent air
contamination by aerosols in each department and at all collection
centres.
 Biohazard bags must be used to dispose of all potentially contaminated
samples-blood tubes, specimen containers, pipettes, pipette tips, reaction
vessels, stoppers etc.

Page 43 of 74
 Safety Manual

 Waste bins are lined with red, yellow and black bags at the start of
each day by trained designated staff.
 All the biohazard bagged bins should be foot pedal operated.

 Waste bins are labelled with logo.

 Leave sufficient area at the top (approximately 1/3 rd part) so that the bag
can be easily closed and secured with an elastic band. It is good
practice to double bag the hazardous waste in case the expected waste
is heavy.
 Remove filled biohazard bags to designated waste areas as frequently
during the day as necessary to avoid build up.
 At the end of each day or after a spill all work surfaces must be
disinfected. i.e. freshly prepared 1% hypochlorite for all work surfaces and
5% phenol for TB lab work surface.

Red Bag: Hand gloves, syringes, any other infected plastic material, blood
tubes and unused needle caps. All the blood sample tubes, urine and stool
containers are discarded in Red bags which are autoclaved and then
given to biomedical waste agency.

Yellow Bag: Contaminated cotton, gauze pieces, human body parts from
histopathology department, microbiology waste, Expired Medicines etc.
All the Microbiology waste are autoclaved and then given to biomedical
waste agency.

Black Bag: Dry waste, papers, dust, wrappers etc.

Blue bag: Intact unbroken empty glass bottles.

c) Sharp Disposal
Sharp containers: Rigid, non-breakable, closed lid plastic containers

with universal biohazard labels are used for sharp disposal at all
processing labs & collection centers.

Sharp Disposal:
1) All needles & broken glassware’s (slide or tubes) are discarded in to sharp
containers.
2) After each vacutainer blood collection needles are directly (without
cutting) discarded into sharp container with the help of push button
holders.
3) After each non-vacutainer blood collection, needles are first cut in
needle cutter & then discarded.
4) Butterfly needle will be directly discarded in sharp container.
5) Once sharp containers are filled 2/3rd the entire container is handed
over to biomedical waste.

Page 44 of 74
 Safety Manual

6) Small capacity of sharp container has to be procured and same has to

be discarded within 48 hours as per BMC guidelines.

Note:
1) Transfer of sharp into another container is not recommended.
2) Use of 1% Hypochlorite in sharp container is not recommended.

All biomedical waste bags & sharp containers are handed over to SMS
Envoclean (Mumbai) or Envirovigil (Thane) or PASSCO (Pune) for further
incineration.

Biomedical waste pick-up frequency: The minimal frequency for

biomedical waste pick-up by biomedical service provider should be
 All Processing labs: Every day
 Collection centres : Alternate days

Records of Biomedical waste: All Franchise, collection centres &

processing labs should have records of biomedical waste collection either
in format provided by service provider or in XXXXX FF.

Reference:
MPCB, Waste management, rules 2016, 28th march 2016.

d) Autoclave procedure
Purpose:
 Autoclave operate at high temperature and pressure in order to kill
microorganisms and spores. They are used to decontaminate certain
biological waste and sterilize media, instruments and lab ware.

Process to be followed:

Page 45 of 74
 Safety Manual

 All the blood sample tubes, urine and stool containers are discarded in
Red bags which are autoclaved for 60 minutes and then given to
biomedical waste agency.
 All Microbiological infectious waste should be discarded in the yellow
bag which are autoclaved for 60 minutes.
 Place all autoclaving material inside the steel mesh tray of autoclave.
Don’t place things directly on the chamber of autoclave.
 All reusable material such as culture plates should be autoclaved in
the discard autoclave steel mesh for 121 degree C at 15 psi for 60
minutes before washing.

Reference:
Refer to the autoclave Instrument SOP for other details

12. Microbiology safety

a) Hazardous Procedure
A number of procedures entail a particular risk of infection. Use of sharps
e.g. scalpels, needles, syringes and breakable glassware should be avoided as far
as possible.

• Syringe and needle: The operator may puncture his skin with the syringe
needle (needle stick injury) during use or disassembly, or another person may

Page 46 of 74
 Safety Manual

accidently puncture himself with an improperly disposed used needle.

Aerosol may be liberated from a vibrating needle on withdrawal from a
vein or culture. Splashing or spraying may be caused by the forceful
ejection of contents.
• Inoculating loop: Vibration of an inoculating loop or straight wire especially
of more than 4 cm long may cause splashing and aerosol production.
Flaming of a wet loop or cooling a hot loop in an agar plate, mixing a slide
agglutination test or spreading a film may produce aerosol.
• Petri dishes: Water of condensation on the agar or in the lid may be
contaminated and spill onto fingers or bench
• Shaking or mixing: Shaking produces an aerosol, even in a closed
container and the aerosol maybe be released on opening the container
soon afterwards. Gross contamination can occur from spillage or breakage
• Centrifugation: Vibration can generate aerosol within the container. Careless
loading or unloading , breakage during centrifuging, or premature opening
after breakage can lead to gross dissemination
• Microbiological safety cabinets: Unless properly installed, maintained
and used, these cabinets can release infection into the air of the room or
exhaust duct. Careless use may cause contamination of the surfaces within
the cabinets, including the hand of users.
• Disposal: Contamination and injury may occur during the collection and
decontamination of discarded cultures, specimen containers and used
equipment especially sharps.

b) Laboratory Biosafety Levels

Four biosafety levels have been recommended based on the infectiousness of
the agent/s.
Biosafety Level-1 (BSL-1): Adherence to standard microbiological practices. No
special requirement as regards containment equipment.
Biosafety Level-2 (BSL-2): In addition to the use of standard microbiological
practice, laboratory coats, decontamination of infectious wastes, limited access,
protective gloves and display of biohazard sign and partial containment equipment
are the requirements for this level.
Most peripheral and intermediate laboratories need BSL-1 or BSL-2 laboratory
facilities.
Biosafety Level-3 (BSL-3): In addition to BSL-2, it has special laboratory clothing,
controlled access to laboratory and partial containment equipment.
Biosafety Level-4 (BSL-4): BSL-3 plus entrance through change room where
laboratory clothing is put on, shower on exit, all wastes are decontaminated before
exit from the facility. It requires maximum containment equipment.

Page 47 of 74
 Safety Manual

Guidelines for Surveillance of lab staff handling microorganisms at Biosafety Level-1

(BSL-1): Historical evidence indicates that the microorganisms handled at this level
are unlikely to cause human disease or animal disease of veterinary importance.
Ideally, however, all laboratory workers should undergo a pre-employment health
check at which their medical history is recorded.
Guidelines for the surveillance of laboratory workers handling microorganisms at
Biosafety Level 2: A pre-employment health check is necessary. The person’s
medical history should be recorded and a targeted occupational health
assessment performed. Records of illness should be kept by the laboratory
management. Women of childbearing age should be made aware of the risk to
an unborn child of occupational exposure to certain microorganisms, e.g. Rubella
virus. The precise steps taken to protect the foetus will vary, depending on the
microorganisms to which the women may be exposed.
Laboratory Facilities In BSL-2
 Laboratory should be designed in such a way that it can be easily cleaned.
 Laboratory contains a sink for washing.
 Laboratory tops are impervious to water but resistant to acids, alkalies and
organic solvents.
 An autoclave to decontaminate infectious material is available.
 Illumination is adequate for all laboratory activities.
 Storage space is adequate.

Guidelines for the surveillance of laboratory workers handling microorganisms at

Biosafety Level 3: Mycobacterium tuberculosis complex culture & DST, Dimorphic
fungi and viruses like JEV, Influenza SARS are examples of agents requiring
biosafety level 3 practices and containment. BSL-3 practices, containment
equipment, and facilities are required for laboratory activities in the propagation
and manipulation of cultures and testing of specimens.
Semi-annual skin testing with purified protein derivative (PPD) of previously skin-test-
negative personnel used as a surveillance procedure.Chest X-Ray, Annual
vaccination with the currently licensed influenza vaccine.
Laboratory Facilities In BSL-3
 Medical examination of all laboratory personnel who work in containment
laboratories – Biosafety Level 3 is mandatory
 Medical contact card for staff
 Standard personal protective equipment must be worn, and respirators might
be required
 Solid-front wraparound gowns, scrub suits or coveralls are often required
 All work with microbes must be performed within an appropriate BSC
 Access hands-free sink and eyewash are available near the exit
 Sustained directional airflow to draw air into the laboratory from clean areas
towards potentially contaminated areas (Exhaust air cannot be re-circulated)
 A self closing set of locking doors with access away from general building
corridors

Page 48 of 74
 Safety Manual

 Access to a BSL-3 laboratory is restricted and controlled at all times.

c) Preventive Measures against Laboratory Infections

These are aimed to protect workers, patients and cultures. Following steps are
to be followed:
 Perform adequate sterilization before washing or disposing waste.
 Provide safety hood.
 Ensure that tissues are handled and disposed off properly.
 Promote regular hand washing and cleaning of bench tops.
 Ensure use of gloves.
 Provide mechanical pipetting devices.
 Protect patients from laboratory personnel with skin or upper respiratory tract
infections.
 Provide special disposal containers for needles and lancets.

PIPETTING
Pipetting and suctioning have been identified as the significant and consistent
causes of occupational infections. Various important precautions that must be
taken while pipetting are:
 Develop pipetting techniques that reduce the potential for creating aerosols.
 Avoid rapid mixing of liquids by alternate suction and expulsion.
 Do not forcibly expel material from a pipette.
 Do not bubble air through liquids with a pipette.
 Prefer pipettes that do not require expulsion of last drop of liquid.
 Drop material having pathogenic organisms as close as possible to the fluid or
agar level.
 Place contaminated pipettes in a container having suitable disinfectant for
complete immersion.
 A variety of pipettes are available. Selection should depend upon the ease
of operation and the type of work to be performed.

OPENING CONTAINERS:
The opening of vials, flasks, Petri dishes, culture tubes, and other containers of
potentially infectious materials poses potential but subtle risks of creating droplets,
aerosols or contamination of the skin or the immediate work area. The most
common opening activity in most health care laboratories is the removal of
stoppers from containers of clinical materials. It is imperative that specimens should
be received and opened only by personnel who are knowledgeable about
occupational infection risks. Various precautions that can be taken in this regard
are:

Page 49 of 74
 Safety Manual

 Open containers with clinical specimens in well-lighted and designated areas

only.
 Wear a laboratory coat and suitable gloves.
 If possible, use an absorbent paper towel to facilitate clean-up and reduce
generation of aerosols

 Specimens which are leaking or broken may be opened only in safety

cabinets. Tubes containing bacterial cultures should be handled with care.
Vigorous shaking of liquid cultures creates a heavy aerosol. When a sealed
ampoule containing a lyophilized or liquid culture is opened, an aerosol
may be created. Ampoules should be opened in a safety cabinet.

LABORATORY ACCESS

 As far as possible children and pregnant women visitors should not enter the
microbiological laboratory.
 Microbiology laboratory has a signage of” restricted entry.”
 Appropriate signs should be located at points of access to laboratory areas
directing all visitors to a receptionist or receiving office for access procedures.

 The universal biohazard symbol shall be displayed at specific

laboratories in which manipulations of organisms with moderate and heavy
risk are being carried out. Only authorized visitors shall enter the laboratory
showing universal biohazard sign. Doors displaying biohazard symbol shall
not be propped open, but shall remain closed when in use.

CLOTHING
 All employees and visitors in microbiological laboratories shall wear
laboratory clothing and laboratory shoes.
 Disposable gloves shall be worn wherever chemical, carcinogenic materials risk
is handled.
 Laboratory clothing including shoes shall not be worn outside the work area.

ACCIDENTS IN LABORATORY
In the microbiological laboratory, bacterial infections pose the most frequent
risk. The important diseases/organisms are:
1) Hepatitis B virus
2) Shigella spp.
3) HIV
4) Salmonella spp. including S typhi
5) Brucella spp.
6) Bacillus anthracis
7) Leptospires,

Version No/Date Page 50 of 74

XX
All rights reserved. Not to be reproduced without authorization
 Safety Manual

8) Yersinia pestis
9) Mycobacteria spp.
10)Histoplasma

MANAGEMENT OF LABORATORY ACCIDENTS

An adequately equipped first-aid box should be kept in the laboratory in a place
that is known and accessible to all members of staff. The box must be clearly
marked and preferably be made of metal or plastic to prevent from damage by
pests.

ENVIRONMENTAL CONTROL:
• The entry to lab is restricted to trained laboratory personnel.
• A entry is made in the register for the visitors before entering the laboratory.
Separate Light Brown colour lab coat is provided to all the visitors as a PPE.
• Biosafety cabinets, ducted to the outside while switched ON would
maintain an inward air flow into the cabinet.
• Access to the culture and DST room is via an anteroom
• Autoclave is provided within the laboratory facility.
• Biological safety cabinet class II A ducted to outside ,is provided
• Centrifuge with aerosol seal buckets is provided.

PERSONAL PROTECTIVE MEASURES

Following protection measures to be followed in the lab by the staff

• All personnel working in culture lab need to wear separate clothing and
separate foot wear.
• While performing culture and DST activities and while reading and
recording results, lab personnel should exercise maximum caution. N95
masks should be worn.
• Remove lab coat before leaving the lab.

d) Biosafety Precautions in TB lab

Definition: Safety to the personnel and environment needed while handling
biological infectious substances such as tubercle bacilli

N-95 respirator: A disposable N 95 mask that has the ability to filter out 95%
of particles greater than 0.3 microns in diameter.

TB lab has all the major facility requirements for handling Mycobacterium
tuberculosis safely and involves minimal risk to the lab personnel if they take
proper precautions and employ proper techniques. Use of the lab is limited to
trained lab personnel.

Page 51 of 74
 Safety Manual

Dos:
• Ensure that lab floor is cleaned with disinfectant every day. The same mop
should not be used for mopping outside the lab bench surfaces.
• Ensure that the instrument surfaces are cleaned with 5% phenol solution
regularly.
• Use double pairs of gloves while working inside bio safety cabinets.
Discard the outer pair of gloves into red bag. Apply disinfectant to hands
before removing the inner pair of gloves.
• Ensure that infectious material is placed away from regular reagents.
• Wash hands thoroughly with disinfectant and tap water before starting work,
after work and before leaving the lab.

Don’ts:
• Eating, drinking smoking, applying cosmetics, use of mobile phones or
applying contact lens in the TB lab.
• Don’t allow unauthorised personnel to enter the TB lab
• Mouth pipetting is prohibited.
• Crowding of lab and bio safety cabinet with material that is not required
inside.

ENTRY AND EXIT REQUIREMENTS FOR CULTURE AND DST FACILITIES

• Enter the anteroom, change the lab coat to the one required that is placed
inside
• Change the street shoes to the lab shoes
• Firmly close the outer door of anteroom
• Open the inner door and make an entry
• Close the door firmly behind you
• The same procedures are followed in reverse order while exiting the facility.

Biological Safety Cabinets:

 All procedures requiring manipulation of TB cultures must be done within
the Bio safety cabinet.
• Switch ON the safety cabinets for at least 15 minutes before use. Note
that the reading on the pressure gauge is satisfactory
• Wear double pair of gloves every time you work inside the cabinet
• Biosafety cabinets need to be cleaned with 5%phenol before work

• Keep containers with 5% phenol inside the cabinet with biohazard logo.
• Arrange all uninfected material towards the right side
• All processed samples on the left.
• Don’t process more than 8 specimens at a time inside the cabinet

Page 52 of 74
 Safety Manual

• After completion of work, wipe all the surfaces with 5% phenol and
discard the wipes in red bags
• Discard the outer glove inside the biosafety cabinet
• Wipe off the inner glove with disinfectant before touching anything else in the
lab

WASTE DISPOSAL AND HANDLING

 All infectious waste should be discarded in the yellow bag
 The waste should then be sterilised at 121oC at 15 psi for 60 mins in the
discard autoclave.
 Place all autoclaving material inside the steel mesh tray of autoclave.
Don’t place things directly on the chamber of autoclave.
 All reusable autoclavable material such as glass ware and petriplates
should be autoclaved in the autoclave steel mesh for 121oC at 15 psi for
60 minutes before washing.

ACCIDENTS AND SPILLAGE;

 Any major accident in the lab should be entered in the accident register
along with remedial measures taken before undertaking further work
 Lab personnel who are accidentally exposed to an infectious TB aerosol or
solution should report the incident as soon as possible to the microbiologist/ lab
director. The microbiologist will see that necessary treatment or health
monitoring is organised without delay.

SPILLS INSIDE THE BIOLOGICAL SAFETY CABINET

A biosafety cabinet is designed to contain spills and associated aerosols which
are released during work within the cabinet. A spill of TB material should be
attended to immediately.
Biological spill kit is available in the TB lab.
The contents of the biological spill are as follows:
1. Double pair of gloves
2. N95 mask
3. Phenol -5%
4. Yellow bag
5. Red bag
6. Eye shield

Decontamination of the work zone is done by application of 5% phenol

disinfectant solution along with a thorough wipe down procedure.

Page 53 of 74
 Safety Manual

Formaldehyde gas decontamination may be required to treat inaccessible

sections of the cabinet interior following a spill.
1. All workers using biosafety cabinets should keep absorbent materials and 5%
phenol within the cabinets
2. Alert all people in lab of immediate area of in event of spill
3. Spread 5% phenol soaked wipe immediately while the biosafety cabinet
continues to operate. Wait for 15-20 minutes.
4. Wear double pair of gloves during the decontamination procedure
5. Contain the spill and decontaminate
6. Use tissues to wipe up the spill working from the edges into the centre.
7. Decontaminate equipment with 5% phenol. Items that are not readily or
easily surface decontaminated should be carefully placed in yellow bags
and removed for further treatment (e.g. treatment by autoclaving)
8. Contaminated gloves and clothes and the lab coat (sleeves are most likely to
be contaminated) should be removed and decontaminate by autoclaving
or soaking in decontaminant.

9. Individuals involved in the spill and clean up should remove

protective clothing, wash their hands and face with an appropriate detergent
soap and report to lab in charge.

SPILLS OUTSIDE THE BIOLOGICAL SAFETY CABINET IN THE TB LAB

Spills on equipment like vortex, centrifuge, incubator, and refrigerator, lab benches,
walls or floors.
1. Immediately indicate to all personnel working in the lab and evacuate for 1
hour to allow dissipation of aerosols created by the spill (negative air pressure
system would clear the aerosols)
2. Leave the biological safety cabinet operating inside the TB lab.
3. Close laboratory doors and post warning signs to prevent others from entering
the lab.
4. Thoroughly wash hands and other apparently contaminated areas with
soap and water. Put on clean disposable gloves.
5. If personal clothing is contaminated, remove all outer clothing and place it in
the autoclave. Put on clean garments.
6. Wear N95 mask, fresh lab coat and double pair of gloves and re-enter after 1
hour.
7. Upon returning to the lab to start decontamination, cover the spill area
with paper towels soaked in 5% phenol. Do not pour decontamination
solution directly on the spill in order to avoid additional release of aerosols.
8. Let stand for 30 minutes then wipe up the spill with the soaked paper
towels and place the used towels in the yellow bag.

Page 54 of 74
 Safety Manual

9. Discard used gloves in red bag

10. In case spill is inside the centrifuge bucket/tube, always use the
aerosol containment cups for centrifuging. Always open the centrifuge
buckets inside the biosafety cabinets. Autoclave the buckets.
11. Wash hands and other apparently contaminated areas again with
soap and water.

AEROSOL GENERATING ACTIVITIES WHICH REQUIRE BIOSAFETY MEASURES IN THE

MYCOBACTERIOLOGY LAB

ACTIVITY

1 Preparing specimens for centrifugation and AFB culture

2 Centrifugation of specimens

3 Inoculating cultures from specimen sediment

4 Handling unopened primary tubes

5 Staining smears of material from culture

6 Manipulating cultures (suspension preparation, vortex and

transferring ) of Mycobacterium tuberculosis

7 Transferring large volumes of cultures or suspensions of bacilli

BIOLOGICAL SAFETY PROCEDURES AND BIOSAFETY SIGNAGES

Biosafety level 2 plus biosafety level 3 practices is considered adequate for

working with Mycobacterium tuberculosis cultures and DST.

BIOSAFETY LEVEL 1

BSL 1 standard microbiological practices

1. Access to the laboratory is limited or restricted to the lab personnel.

2. Work surfaces are decontaminated
3. Liquid or solid waste is decontaminated before disposal.
4. Mechanical pipetting devices are used. Mouth pipetting is prohibited
5. Policies for safe handling of sharps are followed. The needle and
syringe should be promptly place in a puncture proof container.

Page 55 of 74
 Safety Manual

6. Eating, drinking, smoking and applying cosmetics are not permitted in the
work area. Food may be stored in cabinets or refrigerators designated
and used for this purpose only.
7. Persons wash their hands after handling materials involving tuberculosis
organisms and before exiting the lab.
8. All procedures are performed carefully to minimise the creation of
splashes or aerosols.
9. A biohazard sign must be posted at the entrance to the lab whenever
infectious agents are present. The sign must include the name of the
agent in use and the name and phone number of the microbiologist
and the senior technician.
10. Gloves should be worn if the skin on the hands is broken or if the rash
is present.
11. Protective eyewear should be worn for conduct of procedure in which
splashes of microorganisms or other hazardous materials is anticipated.
12. The lab is designed so that it can be easily cleaned. Carpets and rugs
in laboratories are not appropriate.
13. Benches tops are impervious to water and resistant to acids, alkalies,
organic solvents and moderate heat.
14. Laboratory furniture is sturdy. Spaces between benches, cabinets and
equipment are accessible for cleaning.
15. Each lab contains a sink for hand washing.

If the lab has windows that open, they are fitted with fly screens

BIOSAFTEY LEVEL 2

BSL2 STANDARD MICROBIOLOGICAL PRACTICES

All procedures for BSL1 standard microbiological practices and BSL2 SPECIAL
PRACTICES

ALL BSL2 Special Practices

1. The microbiologist or the lab director has the final responsibility for assessing
each circumstance and determining who may enter or work in the
laboratory. For example persons who are immune-compromised may be at
increased risk of acquiring infections.
2. Laboratory coats, gowns, or uniforms are worn while in the laboratory. Before
exiting the lab for non-lab areas (e.g. cafeteria) the protective clothing is
removed.
3. All wastes from lab are appropriately decontaminated before disposal.

Page 56 of 74
 Safety Manual

4. Broken glassware is removed by mechanical means such as brush and

dustpan. Broken glassware should be promptly placed in puncture proof

sharps container with logo.

5. Spills and accidents that result in overt exposures to organisms are
immediately reported to the HOD or lab director.
6. A biosafety manual is prepared or adopted. Personnel are advised of
special hazards and are required to read and follow instructions on
practices and procedures.
7. The microbiologist ensures that the lab personnel receive training on potential
hazards associated with the work involved, the necessary precautions to
prevent exposures and the exposure evaluation procedure.
8. Properly maintained biological safety cabinets are used whenever
procedures involving high potential for creating aerosols are conducted.
These may include centrifuging, grinding, blending, and vigorous shaking pre-
mixing, sonic disruption, opening containers of materials whose internal
pressures may be different from ambient pressures. Centrifuge safety cups
are used for all centrifugations involving M.Tuberculosis are opened only in
a biological safety cabinet.
9. A properly maintained biological safety cabinet with a current and annual
certification and is used according to the manufacturer’s instructions.

BSL2 LAB FACILITIES

All BSL1 Laboratory facility requirements AND
1. Provide lockable doors for facilities that house restricted agents
2. Install Biological safety cabinets in such a manner that fluctuations of
the room supply and exhaust air do not cause the biological safety
cabinets to operate outside their parameters for containment. Locate
biological safety cabinets away from doors, from windows that can be
opened, from heavily travelled areas and from potentially disruptive
equipment so as to maintain the biological safety cabinets air flow
parameters for containment.
3. Illumination is adequate for all activities , avoiding reflection and glare that
could impede vision
4. An autoclave for decontamination is available in the premises.
5. Entry to the culture and DST room is through an anteroom.

BIOSAFTEY LEVEL 3

ALL BSL3 Special Practices

1. The laboratory must be separated from the areas that are open to unrestricted

Page 57 of 74
 Safety Manual

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.
2. 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.
3. 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).
4. The laboratory room must be sealable for decontamination. Air-ducting
systems must be constructed to permit gaseous decontamination.
5. Windows must be closed, sealed and break-resistant.
6. A hand-washing station with hands-free controls should be provided near
each exit door.
7. 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 at all times ensure that proper
directional airflow into the laboratory room is maintained.
8. The building ventilation system must be so constructed that air from the
containment laboratory – Biosafety Level 3 is not recirculated to other areas
within the building. Air may be high-efficiency particulate air (HEPA) filtered,
reconditioned and recirculated 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
pressurization of the laboratory. Consideration should be given to the
installation of audible or clearly visible alarms to notify personnel of HVAC
system failure.
9. All HEPA filters must be installed in a manner that permits gaseous
decontamination and testing.
10. Biological safety cabinets should be sited away from walking areas and out of
crosscurrents from doors and ventilation systems.
11. The exhaust air from Class I or Class II biological safety cabinets which
will have been passed through HEPA filters must be discharged in such a
way as

Page 58 of 74
 Safety Manual

to avoid interference with the air balance of the cabinet or the building
exhaust system.
12. An autoclave for the decontamination of contaminated 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 leakproof containers
according to national or international regulations, as appropriate.
13. 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 also be properly protected with
traps and filters.
14. The containment laboratory – Biosafety Level 3 facility design and operational
procedures should be documented.

13. Bioterrorism plan

XXXXX (India) Pvt. Ltd. is committed to providing a safe working environment

and believes employees have a right to know about health hazards associated
with their work. This Bioterrorism response plan includes policies,
procedures, and responsibilities designed to develop in employees an
awareness of potentially hazardous biological agents in the workplace and to
train employees in appropriate, safe working conditions.

Definition
Bioterrorism is defined as the intentional use of biological agents to inflict disease
and/or death on humans, animals or plants. Thus, crop and livestock as well
as human population are considered as possible bioterrorism targets. The
term biological agent applies to a diverse group of microorganisms as well as
toxins of microorganisms.

Emergency Operations Plan

1. Assigns responsibility to organizations and individuals for specific action
2. Sets forth lines of authority to co-ordinate action
3. Describes how people and property will be protected
4. Identifies personnel, equipment and facilities for use during response and
5. Identifies steps to address mitigation concerns during response and recovery
activities
Emergency Support Facilities (ESF) would include the following departments:
1. Transport and communication
2. Search and rescue
3. Mass Care
4. Health services
5. Public works department

Page 59 of 74
 Safety Manual

The efficiency of this inter-disciplinary cooperation will be crucial to contain and

prevent such events. The Public Health and Medical Response Mechanisms
should be discussed under the following sections:
1. Training
2. Surveillance
3. Laboratory Diagnosis
4. Medical Management and
5. Communications

Training:
Medical personnel, laboratory technicians, epidemiologists, public health
officials, and health administrators play a key role. Therefore they need to have
adequate training and education on issues related to Bioterrorism response. The
training component should have the following elements:

1. Recognition of outbreaks and understanding the epidemiologic clues

2. Treatment of casualties and
3. Protection of high-risk people including the health staff
Surveillance
Look out for epidemiologic clues of bio-warfare such as:

1. Unusual number of sick or dying

2. A disease that is unusual for a given geographical area
3. A disease by an uncommon agent or a vector that is not present in the local area
4. Unusual strains or variants of micro-organisms with anti-microbial resistance
patterns different from those prevalent and
5. Intelligence reports of a potential attack, e.g. aerial spraying or claims by
a terrorist network of a release

Laboratory Diagnosis
Clinical Microbiology laboratories should play a key role in the detection and
identification of biological agents. Due care should be taken while collecting
biological specimens from patients and their contacts. Each sample arriving in
the laboratory should be processed as potentially hazardous. From the receipt of
the sample to the identification, each step should have standard protocol.
Universal precautions such as restricted entry and the movement of staff within the
lab area, use of gloves, gowns, masks and trained manpower in carrying out the
tests is essential. Wash with soap and water if there is a direct contact with a
specimen. Appropriate level of confidentiality should be maintained in disclosing the
results of laboratory diagnosis to avoid misuse of data and create panic in the
community.
Notify the Directorate of Health services.

BIOSAFETY
CDC has classified pathogens in to various Biosafety levels

Page 60 of 74
 Safety Manual

Biosafety level-1 involves routinely isolated pathogens with minimum potential

hazard to laboratory personnel and environment.
Biosafety level-2 involves work with specimens that have a low potential for
creating
aerosols.
Biosafety level-3 involves work on agents with a potential for respiratory
transmission and which may cause serious or potentially lethal diseases.
Biosafety level-4 is required to work with dangerous and exotic agents that pose
a high individual risk of aerosol transmitted laboratory infection.

Most of our labs fall into the Biosafety level 1 and 2 categories.

At BSL2, most of the biological agents can be identified except small pox, VHF
which requires BSL4. Therefore; it would be appropriate to have multi-level
laboratory response network (LRN).
Level A laboratories provide early detection of intentional dissemination of
agents. They use clinical data and standard microbiological tests to decide which
specimens should be forwarded to level B. Level B laboratories help to identify
the pathogens and perform their anti-microbial susceptibility testing. Level C
laboratories provide advanced and specialized testing facility and also take part
in evaluation of new tests and reagents. Level D laboratories are agent-specific.

All specimens or isolates transported from one laboratory to another should

carry the biohazard sign. In the present context of anthrax, DRDO, Gwalior has
been designated as the advanced testing facility. Besides the DRDO other
laboratories that can function as referral centres are:
1. NICED, Delhi
2. NIV Pune
3. Enterovirus research centre Mumbai

Medical Management

The first and foremost objective under the situation will be to save human lives and
to restrict the spread of disease. This may involve appropriate quarantine measures.
It is indeed essential that the clinicians, epidemiologists, para-medical and other
staff, that is, the first responders, be protected and offered prophylaxis. A
national stockpile of vaccines and antibiotics should be maintained to meet such
situations. The big hospitals should have bioterrorism response teams in place
for rapid mobilization of resources and to assist state and local health agencies.

Communication

It is highly recommended that a trained knowledgeable person be identified to

deal with the media. Effective communication with the public through the news
media will be essential to limit the terrorists' ability to induce panic and disrupt daily
life. The need of the hour is to develop

Page 61 of 74
 Safety Manual

1. State-of-the-art communication systems that will support disease surveillance

2. Rapid notification and information exchange regarding disease outbreaks
3. Dissemination of diagnostic results and emergency health information and
4. Coordination of emergency response activities.

An effort should be made to answer some of the queries of the general public.

1. ANTHRAX

A. Transmission
Spores of B.anthracis can live in the soil for years and humans can get infected by:
a. Handling products from infected animals leading to skin lesions.
b. Inhalation of spores.
c. Eating undercooked meat of infected animals.
d. Direct person-to-person spread is extremely unlikely.
B. Diagnosis
a. By isolating B.anthracis from respiratory secretions, skin lesions, blood from
suspected cases.
b. By measuring specific antibodies in the blood of persons with suspected disease.
c. By PCR

C. Treatment
CDC recommends Ciprofloxacin 500 mg b.d or Doxycycline 100 mg b.d for 60 days.
In case of inhalation or intestinal anthrax start on IV regimen and then change
to oral therapy as the patient improves. These antibiotics are also recommended
if there is an exposure to the spores in order to prevent infection Human
anthrax vaccine is only available in the U.S.A. It is 93% effective in protecting
against anthrax. Inappropriate use of antibiotics may lead to resistant strains.
Moreover stockpiling may lead to shortages when an actual emergency occurs.

D. Suspicious Mail
a. A letter or envelope that you are not used to getting regularly such as ones
without a known return address.
b. Letters that may have stains on them, or if the content feels like powder inside.
c. If there is excessive postage.

If any such suspicious mail has to be opened, it should be done with masks on.

Do not handle the mail or package suspected of contamination. If you do put it

down. Slip it into a plastic bag or cover it with a towel or a newspaper. Cordon
off the area. Ensure that all personnel who touched the mail wash their hands
with soap and water. If there is any spillage of the powder, pour 0.5%
hypochlorite solution. Dilute glutaraldehyde may also be used. Notify the local
and state health authorities.

Page 62 of 74
 Safety Manual

2. SMALLPOX
Small pox if used as a biological weapon represents a serious threat to civilian
population because of its case-fatality rate of 30% or more among unvaccinated
persons and the absence of specific therapy. A clandestine aerosol release of
smallpox, even if it infected only 50 to 100 persons to produce the first generation
of cases, would rapidly spread expanding by a factor of 10 to 20 times or more
with each generation of cases.

A. Vaccination
WHO stopped vaccination against smallpox in early 1980s. So individuals above 20
yrs. of age are probably not immune, as residual immunity would only last for about
5-10 yrs. To add to our woes is the fact that at present there are no manufacturers
of the vaccine. A limited stock of vaccine is preserved in select centres. US Food
and Drug Administration (FDA) approves smallpox vaccine for use in person in
special high risk categories:
1. Laboratory workers directly involved with smallpox.
2. Military personnel.
B. Treatment
a. Supportive therapy
b. Antibiotics for secondary bacterial infection.
c. No antiviral drug has yet proved effective for the treatment of smallpox.
d. Recently a new drug cidofovir, a nucleoside analogue DNA polymerase inhibitor
may prove useful in useful in preventing smallpox infection if administered Within 1-
2 days after exposure.

C. Decontamination of Articles Infected with Smallpox

a. Variola virus, if released as an aerosol and not exposed to UV light, may persist
as long as 24 hours under favourable conditions
b. All bedding and clothing of smallpox patients should be autoclaved or
laundered in hot water to which bleach has been added
c. Surfaces should be cleaned with hypochlorite and quaternary ammonia
d. Patients who die of small pox should be cremated.

3. PLAGUE

A. Transmission
Plague remains an enzootic infection of rats, squirrels, prairie dogs and other
rodents on every populated continent except Australia. Human plague most
commonly occurs when plague-infected rat fleas bite humans who then develop
bubonic plague. As a prelude to human epidemics, rats frequently die in large
numbers precipitating the movement of the flea population from its natural
reservoir to humans. Although most persons infected by this route develop
bubonic plague, a small minority will develop sepsis with no bubo, a form of
plague termed primary septicaemia plague. Neither Bubonic nor septicaemia
plague spreads from person to person. A small percentage of patients with
bubonic or septicaemia plague develop secondary pneumonic plague and can
spread the disease by respiratory

Page 63 of 74
 Safety Manual

droplets.

Intentional dissemination of plague would most probably occur via an

aerosol, person contracting the disease by this route develops primary pneumonic
plague.

B. Treatment
Historically, the treatment of plague has been Streptomycin.
CDC recommends oral therapy preferably with Doxycycline or Ciprofloxacin.

C. Post Exposure Prophylaxis

a) Asymptomatic person having household, hospital or other close contact with
persons with untreated pneumonic plague should receive post-exposure
antibiotic prophylaxis for 7 days and watch for fever and cough. Close contact is
defined as contact with a patient at less than 2 meters.
b) Doxycycline is the first-choice antibiotic for post exposure prophylaxis.
c) The patient should remain isolated during the first 48 hours of antibiotic therapy
and until clinical improvement occurs.

D. Disinfection
There is no spore form of Y.pestis, so it is far more susceptible to environmental
conditions than sporulating bacteria such as B.anthracis. Y.pestis is very sensitive to
action of sunlight and heating and does not survive long outside the host. In worst-
case scenario, a plague aerosol is estimated to be effective and infectious for as
long as 1 hour. Hospital rooms of patients with pneumonic plague should
receive terminal cleaning. The bodies of patients who have died following
infection with plague should be handled with routine strict precautions.

4. BOTULINUM TOXIN
Botulinum toxin is the most poisonous substance known. A single gram of crystalline
toxin, evenly dispersed and inhaled, would kill more than 1 million people, although
technical factors would make such dissemination difficult.

A. Treatment
Therapy for botulism consists of
1. Supportive care
2. Passive immunization with equine anti-toxin.
3. Optimal use of botulinum anti-toxin requires early suspicion of botulism. Timely
administration of passive neutralizing anti-body will minimize subsequent nerve
damage and severity of the disease but will not reverse existing paralysis.
4. Standard treatment for detoxification such as activated charcoal may be given
before anti-toxin becomes available, but there are no data regarding their
effectiveness in human botulism.

B. Immunization
1. Botulism can be prevented by the presence of neutralizing antibody in the serum.

Page 64 of 74
 Safety Manual

2. Passive immunity can be provided by equine botulinum antitoxin, while

Endogenous (active) immunity can be induced by immunization with botulinum
toxoid.
3. Only laboratory workers at high risk of exposure & military personnel need to be
immunized.
4. Mass immunization is neither feasible nor desirable for reasons that include
scarcity of toxoid, rarity of natural disease and elimination of the potential
therapeutic benefits of medicinal botulinum toxin.
5. Pre-exposure immunization currently is neither recommended nor available to the
general population.
6. Botulinum toxoid induces immunity over several months and so is ineffective
as post exposure prophylaxis

C. Disinfection
Despite its extreme potency, botulinum toxin is easily destroyed.
Heating to internal temperature of 85 degrees Celsius for at least 5 minutes will de-
toxify contaminated food or drink. Persistence of aerosolised botulinum toxin at
a site of deliberate release is determined by atmospheric conditions and particle
size of the aerosol. Extremes of temperature and humidity will degrade the toxin,
while fine aerosols will eventually dissipate into the atmosphere. At a decay
rate of 1
%/minute, substantial inactivation of the toxin occurs by 2 days after aerosolization.
Covering the mouth and nose with clothing confers some protection when
exposure is anticipated. In contrast with mucosal surfaces, intact skin is
impermeable to botulinum toxin. After exposure to the botulinum toxin, clothes and
skin should be washed with soap and water. Contaminated objects or surfaces
should be cleaned with 0.1% hypochlorite solution.

5. TULAREMIA.
A. History and potential as a Biological Weapon
Tularemia was first described as a plague like disease of rodents in 1911 and,
shortly Thereafter, was recognized as a potentially severe and fatal illness in
humans. Tularemia's epidemic potential became apparent in the 1930s and
1940s, when large waterborne outbreaks occurred in Europe and the Soviet
Union and epizootic-associated cases occurred in the United States

Francisella tularensis has long been considered a potential biological weapon. It

was one of a number of agents studied at Japanese germ warfare research units
operating in Manchuria between 1932 and 1945 conducted research to better
understand the pathophysiology of tularemia and to develop vaccines and
antibiotic prophylaxis and treatment regimens. Tularemia is a bacterial zoonosis.
The causative agent of tularemia, Francisella tularensis, is one of the most
infectious pathogenic bacteria known, requiring inoculation or inhalation of as
few as 10 organisms to cause disease. Humans become incidentally infected
through diverse environmental exposures and can develop severe and
sometimes fatal illness but do not transmit infection to others.

Page 65 of 74
 Safety Manual

B. Diagnosis
A weapon using airborne tularemia would likely result 3 to 5 days later in an
outbreak of acute, undifferentiated febrile illness with incipient pneumonia, pleuritis,
and hilar lymphadenopathy. Specific epidemiological, clinical, and
microbiological findings should lead to early suspicion of intentional tularemia in
an alert health system; laboratory confirmation of agent could be delayed.

Without treatment, the clinical course could progress to respiratory failure,

shock, and death.

C. Disinfection
a. Under natural conditions, F tularensis may survive for extended periods in a cold,
moist environment.
b. In circumstances of a laboratory spill or intentional use in which authorities
are concerned about an environmental risk (e.g., inanimate surfaces wet with
material thought to contain F tularensis), decontamination can be achieved by
spraying the suspected contaminant with a 10% bleach solution (1 part household
bleach and 9 parts water).
c. After 10 minutes, a 70% solution of alcohol can be used to further clean the area
and reduce the corrosive action of the bleach.
d. Soap water can be used to flush away less hazardous contaminations.
Persons with direct exposure to powder or liquid aerosols containing F tularensis
should wash body surfaces and clothing with soap water.
e. Standard levels of chlorine in municipal water sources should protect against
waterborne infection
f. Following an urban release, the risk to humans of acquiring tularemia from
infected animals or arthropod bites is considered minimal and could be reduced
by educating the public on simple avoidance of sick or dead animals and on
personal protective measures against biting arthropods.

D. Treatment
a) Prompt treatment with streptomycin, gentamicin, doxycycline, or ciprofloxacin is
Recommended.
b) Prophylactic use of doxycycline or ciprofloxacin may be useful in the early
post exposure period

Personal Protective Equipment Used

If a specimen from suspected cases should come in for testing, the scientific officer
should take due care while processing. From the receipt of specimen to the
identification step, universal precautions should be followed. The specimen
should be processed, as potentially hazardous and preferably at the end of the
day. The testing area should have restricted entry. Proper protective equipment
such as laboratory gown, face shield and N-95 respirator mask should be worn.

Avoid using automated identification methods: In case an isolate is obtained in a

suspected case of biological warfare agents. The organism will not be identified by

Page 66 of 74
 Safety Manual

Automated system of identification and susceptibility.

14. Covid-19 Safety measures

Measures for protecting the staff from exposure to and infection with SARS-CoV-2,
the virus that causes Coronavirus Disease 2019 (COVID-19), depends on exposure
risk. That risk varies based on the type of work being performed, the potential
for interaction (prolonged or otherwise) with people, and contamination of the
work environment. Employers should adopt infection prevention and control
strategies based on a thorough workplace hazard assessment, using appropriate
combinations of administrative controls, safe work practices, and personal
protective equipment (PPE) to prevent exposures.

a. Social Distancing
Maintaining space between yourself and others is a best practice and is one of
the best tools to avoid exposure to the COVID-19 virus. People can spread
the virus without being sick or knowing they are sick, so it is important to
maintain social distance from others whenever possible. Physical distancing
is required to limit exposure to the COVID-19 virus and slow its spread.
 Everyone IN XXXXX has to follow these physical distancing practices:
 Stay at least 3 feet from others (about an arms’ length) at all times.
 All workstations should be oriented to a minimum of 3 feet apart in all
directions.
 Meetings should take place online instead of a conference room. If
you must meet in person, wipe down surfaces, chairs and equipment
after each use, and maintain physical distancing of at least 3 feet.
 Minimize the use of common breakrooms, caffeteria.
 Rearrange furniture in common areas to maintain physical distancing.
 Handshaking and other forms of physical contact are discouraged.
 Supervisors will be expected to ensure employees self-enforce
physical distancing protocols in all areas.

b. Hand Hygiene
Frequent hand washing is one of the most important actions individuals can
take in preventing the spread of COVID-19.
Detailed Hand washing protocols as per WHO guidelines are clearly mentioned
above in 5 b) section of Hand Hygiene.
When to perform hand hygiene
 Before and after using washroom
 Before and after eating /drinking
 After coughing, blowing or sneezing
 After touching biomedical waste

Page 67 of 74
 Safety Manual

 After touching mask or soiled PPE

 Before and after contact with any patient (during phlebotomy, USG,
during payment at the front desk etc.)

c. PPE and Usage

 Additional considerations for PPE
 Laboratory coats must only be worn in designated areas. When not in use,
they should be stored appropriately; they should not be hung on top of
other laboratory coats, or in lockers or hooks with personal items.
 Aprons & Uniforms to be changed DAILY without fail. All aprons have to
be soaked & washed in water that is 60 degrees or above for at least
30 minutes.
 Appropriate personal protective equipment (gloves, gowns, masks and
eye protectors, etc.) is provided and maintained in a sanitary and
reliable condition in all work areas in which whenever blood and body
substances other potentially infectious materials are handled and in
circumstances during which exposure is likely to occur.
 After removing PPE, always wash hands with soap and water, if
available, for at least 40-60 seconds.
 Apart from the PPE mentioned below, all staff should religiously follow:
o Proper social distancing
o Hand hygiene
o Respiratory etiquette

Page 68 of 74
 Safety Manual

 STEPS FOR DONNING PPE:

 STEPS FOR DOFFING (REMOVING) PPE

Page 69 of 74
 Safety Manual

 MASK ETIQUETTE
Cover your mouth and nose with a tissue when you cough or sneeze. If you
do not have tissues, it is recommended that you cough or sneeze into the inside
of your elbow, not into your hands. Throw tissue in the trash and immediately
wash your hands with soap and water or use hand sanitizer.

d. Decontamination
 Cleaning agents and disinfectants:
o Sodium hypochlorite
 1% Sodium Hypochlorite can be used as a disinfectant for
cleaning and disinfection.
- The solution should be prepared fresh.
- Leaving the solution for a contact time of 15 minutes is
recommended.
 4-6 % hypochlorite to decontaminate blood spills
- The solution should be prepared fresh.
- Leaving the solution for a contact time of 15 minutes
is recommended.
o Alcohol (e.g. isopropyl 70% or ethyl alcohol 70%/ Ethanol)
- Can be used to wipe down surfaces where the use of bleach is
not suitable, e.g. metals. Isopropyl alcohol is preferred for
electrical adjacent surfaces.
- The contact time should be at least 5 minutes.
o Detergent Soap
- The contact time should be 10 minutes.

Page 70 of 74
 Safety Manual

- If surfaces are dirty, they should be cleaned using a detergent or

soap and water prior to disinfection.
 Disinfectant or 1% sodium hypochlorite solution should be applied to
surfaces using a damp cloth (not a soaked cloth). They should not be
applied to surfaces using a spray bottle, as coverage is uncertain and
spraying may promote the production of aerosols. The creation of aerosols
caused by splashing liquid during cleaning should be avoided. A steady
sweeping motion should be used when cleaning either floors or horizontal
surfaces, to prevent the creation of aerosols or splashing.
 When cleaning keyboards, light switches and other electrical
equipment ensure that the cloth has NO EXCESS disinfectant. It should be
only a light application.
 Remove curtains/ fabrics/ quilts for washing, preferably using the hot
water cycle. For hot-water laundry cycles, wash with detergent or
disinfectant in water at 70ºC for at least 25 minutes.

S.No. Surface Disinfectant to be used Frequency Person responsible

Door knobs, Door jambs, Front
1 desk counter, Chair arm rests, 1% Sodium hypochlorite Every 3-4 hours Housekeeping staff
table tops

Keyboards, mouse, fan/AC/light

2 70% IPA Every 3-4 hours Housekeeping staff
switches/ telephones

Centrifuges and instrument Housekeeping staff /

3 70% IPA Every 3-4 hours
surfaces Respective technician
Toilet bowls and accessible
4 1% Sodium hypochlorite Every 3-4 hours Housekeeping staff
surfaces of toilet (non-
metal)
70% IPA or detergent
5 Metal surfaces in toilet Every 3-4 hours Housekeeping staff
solution
6 Walls, mirrors, windows 1% Sodium hypochlorite Once daily Housekeeping staff
7 Floors Detergent solution 4 times a day Housekeeping staff
Wipe down with 70% IPA
8 Blinds/curtains Once daily Housekeeping staff
or detergent solution
9 Blood spills 4% Sodium hypochlorite As required Housekeeping staff
Tourniquets, phlebotomy chair After every
10 1% Sodium hypochlorite Phlebotomy staff
arm rests patient
ECG leads, BP Cuff, Stethoscope,
After every Respective non-path
11 USG probes, Spirometry 70% IPA
patient technical staff
equipment
12 Buckets 1% Sodium hypochlorite Prior to filling Housekeeping staff
Visibly dirty/soiled floor, table top,
13 Detergent solution As required Housekeeping staff
wall or any non-electronic surface

 The Inner & Outer surface of bins/Containers/Trolleys to be disinfected using

1% Hypochlorite solution daily.
 Safety Manual

Waste BMW Container Treatment In Lab

PPE – Gloves & Plastic shoe cover/Head
cover
PPE – Plastic Goggles / Face shield
PPE – Gown & Mask
Sample Tubes/Containers
Red Bag No treatment
To be cleaned using 1% Hypochlorite solution
NA
& can be reused.
Soak in 1% Hypochlorite Solution in double
Yellow Bag layered yellow bag. (To ensure adequate
strength & No leakage)
To be Autoclaved as a routine process in NABL
Accredited Labs. Pre-Treat sample in 1%
Red Bag
Hypochlorite for 30 mins and discard in red
bags (All other labs)

 Decontamination protocol for Molecular Diagnostic department(Processing lab)

There is likely hood of carryover contamination of carryover PCR pdts during
high load of COVID 19 RT-PCR processing in Molecular Biology Lab at the
level of extraction, reagent preparation & PCR set up. As not all PCR reagents
contain UNG/UDG, a decontamination protocol was established as follows using
oxidizing agents like Hypochlorite & UV Photoreaction decontamination of post
PCR amplicons. As the hazards & the longer duration time associated with
gaseous decontamination like formaldehyde fumes, the above decontamination
protocol is followed.
 FREQUENCY: Once in 15 days
 HOW DOES PHOTOREACTION WORKS:
UV-Induced Thymine Dimers: In this procedure, ultraviolet light (254 to 300
nm) photocrosslinks pairs of adjacent pyrimidine bases into cyclobutane-
like dimers (Figure 5).The dimers are composed mostly of thymine::thymine
(TT) dimers, although a few thymine::cytosine (TC) and a rare
cytosine::cytosine (CC) combinations are formed as well. Once created,
these modified structures cannot be removed from the dsDNA (or ssDNA)
templates because most of the thermally-stable DNA polymerases
possess little or no exonuclease activity. This contamination control
procedure exploits the presence of the thymine dimers (TT) since they
sterically block extension of the incomplete strand when encountered by the
DNA Polymerase.
 PROCEDURE TO BE FOLLOWED:
The reagent room LAF surfaces, the pipettes, the PCR racks, tube racks & the
picofuge is thoroughly cleaned as follows with the order of hypochlorite
cleaning, followed by mili Q water followed by 70% IPA & then exposed to
UV light. . The order of cleaning is:
a) We tissue papers sufficiently with 1% freshly prepared hypochlorite.
Allowed to react for 10 min with all the surfaces.
b) Then clean with RO water/ miliQ water with wet tissue papers.
c) Then wipe these twice with 70% Isopropyl Alcohol.
The preference of surfaces to clean with above solutions is:
 Safety Manual

Floor by house keeping

Pipettes
Picofuge
Tube racks
LAF/ BSC walls & surfaces
Markers & pens
Note: No metal equipment is cleaned with hypochlorite except the LAF &
BSC surfaces. Also the metallic surfaces once cleaned with hypochlorite,
should be thoroughly wiped with Milq water to remove excess of bleach.
 REFERENCE:
1. Control of Contamination Associated with PCR and Other
Amplification Reactions by Theodore E. Mifflin, Ph.D., DABCC
2. Fumigation with formaldehyde Safety Office, The University of
Nottingham 17th July 2012.

e. Other Safety measures

 Refrain from touching eyes, nose or mouth with gloved or bare hands.
 Refrain from using mobile electronic devices (for example, mobile
telephones, tablets, laptops, flash drives, memory sticks, cameras and/or
other portable devices, including those used for DNA/RNA sequencing)
when not specifically required for the laboratory procedures being
performed.
 Keep mobile electronic devices in areas where they could not easily
become contaminated or act as a fomite for infection. Where close
proximity of such devices to biological agents is unavoidable, ensure they
are either protected by a physical barrier or decontaminated before
leaving the laboratory (Mobile phones to be kept in clear plastic bags).
 Use of Aarogya Setu is continued on best effort basis on compatible
mobile phones. This facilitates timely provision of medical attention to
those individuals who are at risk.(MHA, GOI, No. 40-3/2020-DM-I(A), 23rd
Mar, 2021.
 Aarogya Setu app status (GREEN/RED) is checked for all staff before
checking inn on duty and is updated & maintained by managers in form
of google drive on daily basis.
 Environmental health and safety (EHS) officers are appointed for all the
centres and are responsible for every aspect of the centre compliance
pertaining to COVID-19 complication.
 Staff that have any symptoms related to the virus, a temperature of 99.32°F
or over, fever, dry cough, sore throat or respiratory issues has to self-isolate
and not come to the workplace. Manager and local health authorities are
informed for advice. All personnel follow fundamental controls e.g.
regular handwashing, no physical contact and follow the 1.5 metre
 Safety Manual
physical
 Safety Manual

distance (social distancing) rule to minimize close contact. If any symptoms

develop, personnel must self-isolate.
 If staff/staff relative is tested positive for COVID-19, they must not return to
site/work until they produce a negative test result. Staff confirmed with
COVID-19 is monitored by state health authorities during their isolation
period and follow the guidelines provided.
 Thermal scanning for patients: Infra-red thermometer is used for each of the
walk in patients at the entry point. Acceptable temperature is below 99oF.
 Plastic gloves are provided to all the patients before entering the centres.
 For any external visitors-visiting the labs ,Thermal scanning is done, an entry is
made in the register before entering the laboratory noting down the
Aarogya Setu status, vaccination status and the other details of the
visitor. Visitors are screened and restricted from visiting, regardless of
their vaccination status, if they have: current SARS-CoV-2 infection;
symptoms of COVID-19; or prolonged close contact (within 6 feet of an
infected person for a cumulative total of 15 minutes or more over a 24-
hour period) with someone with SARS-CoV-2 infection in the prior 14
days or have otherwise met criteria for quarantine.(CDC guidelines-27th
April 2021)
 Provision of Hand wash or sanitizer is made at all entry & exit points and
common areas.
 Ensure that no patient uses the water cooler by themselves – put up posters
that if they require water, it will be provided to them in disposable
glasses. Alternatively, one staff can stay near the water cooler to give
patients the water.
 Provide tissues and no-touch receptacles (i.e. waste container with foot
operated lid or uncovered waste container) for used tissue disposal.

REFERENCES
 All India Institute of Medical Sciences (AIIMS, New Delhi). COVID-19
Preparedness Document. 2020.
 WHO Document Ref no: WHO/2019-nCov/IPC_PPE_use/2020.3
 WHO Document Ref no: WHO/2019-nCov/IPC_Masks/2020.3
 WHO Document Ref no: WHO/2019-nCoV/IPC_WASH/2020.2
 WHO Document Ref no: WHO/2019-nCoV/IPC/2020.3
 https://medlineplus.gov/ency/patientinstructions/000452.htm
 https://www.cdc.gov/coronavirus/2019-ncov/hcp/infection-
controlrecommendations.html
 https://www.cdc.gov/coronavirus/2019-ncov/hcp/guidance-risk-
assesmenthcp.html
 CPCB Guidelines & BMW handling