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Laboratory safety


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Laboratory safety

  1. 1. Laboratory Safety Dr Varsha Shahane
  2. 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. 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. 4. Introduction Why Biosafety Practices? Protection:• workers• “products”• co-workers• lab support personnel• environment 2.1
  5. 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. 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. 7. Safety Resources
  8. 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. 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. 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. 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. 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. 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. 14. Post Exposure• Write a report and reasons for accident• Actions taken to avoid future accidents• Training
  15. 15. Chain of Infection Chain of infectionReservoir of pathogen Portal of escape Transmission Route of entry/infectious dose Susceptible host Incubation period
  16. 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. 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. 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. 19. Containment Levels ↔ Risk group of pathogensCONTAINMENT 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. 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. 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. 22. Biosafety Level 1Standard Microbiological Practices 2.3
  23. 23. Biological safety cabinet - class I
  24. 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. 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. 26. Biological safety cabinet - class II
  27. 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. 28. Biosafety Level 2 Laboratory Facilities (Secondary Barriers)• BSL-1 Facilities PLUS: – Autoclave available – Eyewash station available 2.4
  29. 29. Biosafety Level 2 Facility Construction (Secondary Barrier)Requirements: – Location - separated from public areas – Structure - normal construction – Ventilation - directional 2.4
  30. 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. 31. Biosafety Level 2 Special Practices Needles & sharps precautions• DON’T place needles or sharps in office waste containers 2.4
  32. 32. Biosafety Level 2 Special PracticesNeedles and Sharps Precautions (cont.)– DON’T touch broken glass with hands 2.4
  33. 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. 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. 35. Biological safety cabinet – class III
  36. 36. Biosafety Level 3 Safety Equipment (Primary Barriers)• BSL-1 and 2 Safety Equipment PLUS: – Respiratory protection may be indicated 2.5
  37. 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. 38. Biosafety Level 3Laboratory 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. 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. 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. 41. Biological Waste• Types – cultures, stocks, isolates – materials containing or contaminated with blood – sharps – pipettes, wrappers, tips – All materials used in the lab
  42. 42. Specimen Disposal
  43. 43. Decontamination• Sterilization• Disinfection
  44. 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. 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. 46. Decontamination Methods• Heat• Chemical• Radiation
  47. 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. 48. Physical hazards• Fire• Electrical accidents – shock, short circuit• Noise levels• Equipment accident – slipping, breaking, falling• UV light exposure• Compressed gases• Work stress
  49. 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. 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. 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