1. Laboratory Safety
Dr Varsha Shahane

2. Objectives
• To define the essential components of a laboratory
safety program
• To administer the program to meet safety
requirements
• To evaluate the program for regulatory compliance
• To identify hazardous materials and procedures in
the laboratory

3. Hazards in the lab
• Biological : exposure to blood and body fluids and
specimens that harbour HIV, HBV, HCV etc,
bioterrorism)
• Chemical : acids, alkalies, toxic chemicals
• Radiological : ineffective radioactive waste disposal
• Fire
• Electrical accidents

4. Introduction
Why Biosafety Practices?
Protection:
• workers
• “products”
• co-workers
• lab support personnel
• environment
2.1

5. Safety management plan
• Development, implementation and enforcement of a
quality safety management program → WORKER
SAFETY
• Occupational Safety and Health Administration
(OSHA) – risk assessment of each lab task
• Well written ‘Safety manual’ – defines the program
and identifies responsible people
• WHO has published laboratory safety guidelines and
ACDP ( Advisory Committee on Dangerous
pathogens) has formulated biosafety levels for all
microbes

6. Safety Documentation & Records
Laboratory Safety Manual - Policies and Procedures
• Sample Contents:
– Housekeeping
– Personal protection
– Safe decontamination of equipment
– Decontamination & Waste Disposal
– Emergency procedures
 In-lab first aid
 Accidental injury
 Post exposure prophylaxis
 Contacts
– Personnel responsibilities
Hygiene

7. Safety Resources

8. contd
• Provision of necessary time and resources to
produce and implement the plan and communicate it
to employees
• Each task is assessed for the risks and accordingly the
plan is framed :
 standard precautions
 personal protective equipment (gloves, coats, masks)
 engineering and work practice controls, work place design
 vaccination
 safe handling and disposal of hazardous waste
 use of safety devices

9. contd
 blood borne pathogens
 medical surveillance program
 chemical hygiene plan
 infection control procedures
 hazard communications
 showers and eye wash station, safety eye glasses
 record keeping
 waste disposal
 fire safety
 spill cleanup

10. Contd..
• Tie hair, no cosmetic use
• Closed footwear
• No smoking, eating or drinking in lab
• Screening of lab personnel for contagious and
infectious diseases

11. Contd..
• SAFETY OFFICER & SAFETY COMMITTEE – safety
policies documented in safety manual, check for
compliance, training of employees, safety audits and
risk assessment, maintain records and carry out
inspections. Ensure that the current legal
requirements and GLP standards are followed.

12. Action Plan for Implementing Safety Practices
• Identify hazards
• Assess level of risks
– Prioritize risk
• Establish and implement safety polices and
procedures
• Conduct safety specific training
– Must be a priority
– Communication is key
• Perform regular audits and assessments

13. In Case of Exposure
• Be ready for the emergency before hand
- Familiar with exposure specific policies
- Conduct drills
- Keep post exposure medicines available
- Check periodically for stock and expiry of
medicines
• Report immediately
• Go to the nearest, first available doctor

14. Post Exposure
• Write a report and reasons for accident
• Actions taken to avoid future accidents
• Training

15. Chain of Infection
Chain of infection
Reservoir of pathogen
Portal of escape
Transmission
Route of entry/infectious dose
Susceptible host
Incubation period

16. Laboratory hazards - Factors
Factors determining the impact of hazards:
• Exposure levels
• Duration of exposure
• Toxicity or pathogenecity of the hazardous material
• Safety controls
• Number and types of contact with the hazard
• Host factors – age, health etc

17. Biological hazards
• Infectious agents transmitted through body
secretions or tissue
• HIV, Hepatitis B and C, Mycobacterium tuberculosis
etc
• Safe work practices, well designed labs, containment
equipment, management controls, prevention of
unauthorized entry, removal of bioterrorism agents

18. Risk Group of Characterization
pathogens
1 unlikely to cause disease. Not considered infectious
2 moderate individual and low community risk. Unlikely
to cause serious disease or be transmitted. Prophylaxis
and treatment available(Enterobacteriaceae,HSV, Candida,
Mycobacterium avium)
3 High individual and community risk. Causes serious
infections, but not readily transmitted. Effective
treatment and prevention available (Mycobacterium
tuberculosis, Bacillus antracis, Brucella,S.typhi. Shigella,Rabies virus,
Yellow fever virus, Hepatitis B virus,West Nile virus)
4 High individual and community risk. Causes serious
infections, readily transmitted. Effective treatment and
prevention not available (Ebola and Marburg virus, Variola, KFD

19. Containment Levels ↔ Risk group of pathogens
CONTAINMENT LEVEL REQUIREMENTS
Lab to be cleaned daily and have washing facilities. No
1 eating/drinking/smoking/cosmetics/mouth pipetting in lab
Above measures + limited access, daily cleaning of benches,
lab size 24 m3/worker. Personal protective clothing,
2 biosafety cabinet class I and autoclave for sterilization of
waste must be available
Above 2 + lab should be away from populated area, have a
3 biohazard sign on it,fully well equipped, air flow monitored
and HEPA filtered, class III biosafety cabinet required.
Sealed class III biosafety cabinets, sophisticated air
4 movement control and filtration, all safety features.

20. Designing for safety
 Facility design
 Water supply/sinks for hand washing
 Ventilation
 Personal protective equipment (PPE)
 Biosafety cabinets – for product, personal
and environmental protection

21. Biosafety Level 1
Standard Microbiological Practices
• Restrict or limit access when
working
• Prohibit eating, drinking and
smoking in the laboratory
• Pipetting by mouth strictly
forbidden
2.3

22. Biosafety Level 1
Standard Microbiological Practices
2.3

23. Biological safety cabinet - class I

24. Standard practices also include:
• Keep work areas uncluttered and clean
• No food in lab refrigerator
• Minimize splashes and aerosols
• Decontaminate work surfaces daily
• Maintain insect & rodent control program

25. Biosafety Level 2
Safety Equipment (Primary Barriers)
In addition to BSL-1:
• Use biosafety cabinets (class II) for work with
infectious agents involving:
– Aerosols and splashes
– Large volumes
– High concentrations
2.4

26. Biological safety cabinet - class II

27. Biosafety Level 2
Facility Design (Secondary Barriers)
Requirements:
 Laboratories have lockable doors
 Sink for hand washing
 Work surfaces easily cleaned
 Bench tops are impervious to water
 Sturdy furniture
2.4

28. Biosafety Level 2
Laboratory Facilities (Secondary Barriers)
• BSL-1 Facilities PLUS:
– Autoclave available
– Eyewash station
available
2.4

29. Biosafety Level 2
Facility Construction (Secondary Barrier)
Requirements:
– Location - separated from public areas
– Structure - normal construction
– Ventilation - directional
2.4

30. Biosafety Level 2
Special Practices
Needles & Sharps Precautions
– Use sharps containers
– DON’T break, bend, re-sheath or reuse
syringes or needles
2.4

31. Biosafety Level 2
Special Practices
Needles & sharps precautions
• DON’T place needles or sharps in office waste
containers
2.4

32. Biosafety Level 2
Special Practices
Needles and Sharps Precautions (cont.)
– DON’T touch broken glass with hands
2.4

33. Biosafety Level 2
Special Practices
• Identify “clean” and “contaminated” areas
– Use appropriate warning signs
• Decontaminate work surfaces
• Report spills and accidents
• Remove gloves, lab coats before leaving work
area
• No animals in laboratories
2.4

34. Biosafety Level 3
Safety Equipment (Primary Barriers)
• BSL-1 and 2 Safety Equipment PLUS:
– BSC class
II or III to
manipulate
infectious
material
2.5

35. Biological safety cabinet – class III

36. Biosafety Level 3
Safety Equipment (Primary Barriers)
• BSL-1 and 2 Safety Equipment PLUS:
– Respiratory protection may be indicated
2.5

37. Biosafety Level 3
Laboratory Facilities (Secondary Barriers)
• BSL-1 and 2 Facilities PLUS:
– Separate building or isolated zone
– Double door entry
– Directional inward airflow
– Single-pass air; 10-12 air changes/hour
2.5

38. Biosafety Level 3
Laboratory Facilities (Secondary Barriers)
• BSL-1 and 2 Facilities PLUS (contd):
– Enclosures for aerosol generating equipment
– Room penetrations sealed
– Walls, floors and ceilings are water resistant for
easy cleaning
2.5

39. Biosafety Level 3
Special Practices
• BSL-2 Special Practices PLUS:
– Work in certified BSC
– Use bioaerosol-
containing equipment
– Decontaminate spills
promptly
2.5

40. Biosafety Level 4 – Maximum Containment
• BSL -3 practices plus:
– Clothing change before entering laboratory
– Shower on exit
– All materials decontaminated on exit from facility
• Safety Equipment:
– Class III Biosafety cabinet
– Class I or II biosafety cabinet
– with full-body, air supplied, positive personnel suit

41. Biological Waste
• Types
– cultures, stocks, isolates
– materials containing or contaminated with blood
– sharps
– pipettes, wrappers, tips
– All materials used in the lab

42. Specimen Disposal

43. Decontamination
• Sterilization
• Disinfection

44. Decontamination
Chemical
• General Lab Use - Hypochlorite Solutions
– Large Spills/Large Organic Load
• undiluted from bottle
– Small Spills/Virus Inactivation
• 10% - 1:9
– General Surface Disinfection
• 1% - 1:99

45. Contd....
• Handling specimens and spills
– All specimens should be considered as potentially
dangerous
– Specimens should be in well sealed, leak proof, solid
containers
– Wear gloves and cover abrasions/cuts on hands
– Requisition forms that are blood stained should be
rejected and handled with gloved hands
– Wash hands thoroughly several times in a day with soap
and water
– Spills : Place a newspaper on the spill. After it gets soaked,
pour 1% -5% hypochlorite on it and leave it for 10 minutes.
Then mop the area with disinfectant

46. Decontamination
Methods
• Heat
• Chemical
• Radiation

47. Chemical hazards
• Label lab chemicals with their hazard classification –
irritant, corrosive, flammable, carcinogen etc
• Store reactive chemicals seperately ( acids x bases,
oxidizers x reducing agents). Flammables are stored
in flamable safe cabinets
• Use of carcinogens / toxic chemicals to be used only
by trained personnel and in designated area
• Chemical hygiene plan, engineering and work
practice controls, exposure monitoring and waste
management

48. Physical hazards
• Fire
• Electrical accidents – shock, short circuit
• Noise levels
• Equipment accident – slipping, breaking, falling
• UV light exposure
• Compressed gases
• Work stress

49. Radiation hazards
• Type of radiation – α, β or ϒ, quantity and source of
exposure (internal /external)
• Time, distance, shielding – engineering and work
practice controls
(designated areas, lead aprons, monitoring badges)

50. Routine safety precautions
• Centrifugation
– Check tubes/vials for cracks
– Properly balanced
– Wait for a complete stop
– In case of breakage, clean the buckets with disinfectant
– For infectious agents, seal the tubes

51. Contd....
• Electrical safety
– Must know master switches and circuit breaker boards
– Plugs/ cords in worn or broken conditions X don’t use
– All electrical equipment must be grounded
– All shocks to be investigated
• Corridor cautions
– Swinging doors ; should open into corridors
– Use only one side of corridor for storage of equipment
– Look for spills, glass etc on the floors
– Always walk, never run