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Laboratory safety

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LABORATORY
SAFETY
• Although it is the legal responsibility of hospital and laboratory
managers to provide for a safe working environment
• One person in the laboratory should be designated the safety
officer, whose duties are to see that safety standards and
guidelines are written and published, that employees are
informed of these standards through regularly scheduled
laboratory courses and in service briefings, and that a system
is in place to monitor compliance.
BIOSAFETY: Preventing lab-
acquired infections
• Bacteria
• Viruses
• Fungi
• Human blood, unfixed tissue
• Human cell lines
Why is Biosafety Important
• Laboratories recognize hazards of processing infectious agents
• Guidelines developed to protect workers in microbiological
and medical labs through engineering controls, management
policies, work practices
GENERALSAFETYRULESAND
REGULATIONS
• Whenever a person enters a laboratory he/she should remove
the footwear out of the laboratory.
• Dyes and chemicals are used in the laboratory due to which
our skin can get harmed as well as it can damage our clothes
so laboratory gown or apron should be used compulsory.
• Instruction given by the instruction should be noted and
followed carefully.
• Eating and drinking is not allowed in laboratory.
• Do not put anything in your mouth like pen, pencils, and
fingers etc.
• Hair should be packed with hair cap to avoid falling of hair on
face, working place as well as burning Bunsen burner

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Dr. Ahmed Samhanد حاتم البيطاررر اكاديمية زويل .pdf
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Dr. Ahmed Samhanد حاتم البيطاررر اكاديمية زويل .pdf
 

Laboratory safety

  • 2. • Although it is the legal responsibility of hospital and laboratory managers to provide for a safe working environment • One person in the laboratory should be designated the safety officer, whose duties are to see that safety standards and guidelines are written and published, that employees are informed of these standards through regularly scheduled laboratory courses and in service briefings, and that a system is in place to monitor compliance.
  • 3. BIOSAFETY: Preventing lab- acquired infections • Bacteria • Viruses • Fungi • Human blood, unfixed tissue • Human cell lines
  • 4. Why is Biosafety Important • Laboratories recognize hazards of processing infectious agents • Guidelines developed to protect workers in microbiological and medical labs through engineering controls, management policies, work practices
  • 5. GENERALSAFETYRULESAND REGULATIONS • Whenever a person enters a laboratory he/she should remove the footwear out of the laboratory. • Dyes and chemicals are used in the laboratory due to which our skin can get harmed as well as it can damage our clothes so laboratory gown or apron should be used compulsory. • Instruction given by the instruction should be noted and followed carefully.
  • 6. • Eating and drinking is not allowed in laboratory. • Do not put anything in your mouth like pen, pencils, and fingers etc. • Hair should be packed with hair cap to avoid falling of hair on face, working place as well as burning Bunsen burner
  • 7. • Laboratory doors and windows should be kept closed during experiments are going on. • Extra material like bags ,cloths, purses and books should be kept at proper places like drawers or shelf’s.
  • 8. • The working area should be kept free from all these extra material. • Before starting your work on your work bench the bench top should be disinfected by ethanol, phenol,Lysol or cetylpyridinium chloride before and after each laboratory work. • Hands should be washed properly before and after each task.
  • 9. • These above precaution may lessen chances of infection or contamination due to cultures. • Smoking is completely prohibited in microbial laboratory. • Microbial cultures should be handled properly and should not be taken out of laboratory.
  • 10. • Mouth pipetting should be avoided and mechanical pipetting devices should be used. • All laboratory work should be carried out carefully to avoid aerosol formation. • Talking ,laughing should be avoided because it can result into contamination. • All contaminated material should placed into a proper container.
  • 11. • Culture plates, tubes should be discarded in basket. • All microbial cultures should be either incubated or refrigerated, Cultures should not be kept in drawers, cupboard or desk. • Pipettes, glass rods should be properly disinfected or sterilized after each use. • Before and after each use nichrome wireloop should be sterilized and then used or kept.
  • 12. • All laboratory instruments should be operated as per standard operating procedure. • Bunsen burner are very dangerous so the should be handled with care and should be turned of when not in use. • All electrical appliances such as oven, autoclave, centrifuge and other such devices should be immediately turned off after each use.
  • 13. • All accidents, injuries or broken glassware should be reported immediately. • Alcohol should be kept away from flame. • Waste paper, threads, broken glassware or used pH strips should be discarded safely. • The experiments that are permitted should only carried out. • Keep the laboratory clean.
  • 14. Biosafety Levels • Precautions so people researching or trying to identify organisms do not become infected • While handling or testing clinical specimens, workers could accidentally infect themselves or coworkers • Labs must adhere to very specific safety regulations to work with organisms that pose a threat to human health
  • 15. Laboratories divided on basis of nature of microbes • Labs divided into 4 biosafety levels; protective practices increase with each • Biosafety Level 1 labs - work with least dangerous agents, require fewest precautions • Biosafety Level 4 labs - have strictest methods because dealing with agents that are most dangerous to human health
  • 16. Biosafety Level 1 Practices • safety equipment and facilities appropriate for work with defined and characterized strains of viable micro organisms not known to cause disease in healthy adult humans. • The laboratory is not necessarily separated from the general traffic patterns in the building. • Work is generally conducted on open bench tops using standard microbiological practices
  • 17. • Special containment equipment or facility design is neither required nor generally used. • Laboratory personnel have specific training in the procedures conducted in the laboratory and are supervised by a scientist with general training in microbiology or a related science. • A biological safety cabinet is generally not required for work involving these agents. • Examples: Bacillus subtilis, Naegleria gruberi, Infectious canine, hepatitis virus, E.coli
  • 18. Biosafety Level 2 Practices • safety equipment and facilities appropriate for work done with a broad spectrum of indigenous moderate-risk agents present in the community and associated with human disease in varying severity. It differs from biosafety level 1 in that: a) laboratory personnel have specific training in handling pathogenic agents and are directed by competent scientists; b) access to the laboratory is limited when work is being conducted;
  • 19. • c) extreme precautions are taken with contaminated sharp items; • d) certain procedures in which infectious aerosols or splashes may be created are conducted in biological safety cabinets or other physical containment equipment. • A Class I or Class II biological safety cabinet is highly recommended for work involving these agents • Examples : measles virus, salmonellae, Toxoplasma spp, Hepatitis B Virus.
  • 20. Biosafety Level 3 Practices • safety equipment and facilities appropriate for work done with indigenous or exotic agents with a potential for respiratory transmission which may cause serious and potentially lethal infection. • More emphasis is placed on primary and secondary barriers to protect personnel in the contagious area, the community, and the environment from exposure to potentially infectious aerosols. • A Class I or Class II biological safety cabinet is required for work involving these agents. • examples: M. Tuberculosis, St. louis encephalitis virus, Coxiella Burnetii, Bacillus anthracis (production level)
  • 21. Biosafety Level 4 Practices • safety equipment and facilities appropriate for work done with dangerous and exotic agents which pose a high risk of life threatening disease. • May be transmitted via the aerosol route, and for which there is no available vaccine or therapy. Members of the laboratory staff have specific and thorough training in handling extremely hazardous infectious agents and they understand the primary and secondary containment functions of the standard and special practices, the containment equipment, and the laboratory design characteristics
  • 22. • . They are supervised by competent scientists who are trained and experienced in working with these agents. Access to the laboratory is strictly controlled by the laboratory director. • The facility is either in a separate building or in a controlled area within a building, which is completely isolated from all other areas of the building. • A specific facility operations manual is prepared or adopted. • A Class III biological safety cabinet is required for work involving these agents. • Examples: Ebola zaire, Sin Nombre Virus, Rift Valley Fever
  • 23. BIO SAFETY CABINETS • The terms biological safety cabinet and biosafety cabinet have been widely used to describe a variety of containment devices equipped with HEPA filter(s), designed to provide personnel or both personnel and product protection from biohazardous materials.
  • 25. class I Biological safety cabinets A. front opening B. sash C. exhaust HEPA D. exhaust plenum
  • 26. • A stream of inward air moving into the cabinet contains aerosols generated during microbiological manipulations. It then passes through a filtration system that traps all airborne particles and contaminants. Finally, clean, decontaminated air is exhausted from the cabinet. • The filtration system usually consists of a pre-filter and a HEPA (High Efficiency Particulate Air) filter.
  • 27. • Although the Class I cabinet protects the operator and the environment from exposure to biohazards, it does not prevent samples being handled in the cabinet from coming into contact with airborne contaminants that may be present in room air. • All Class I biological safety cabinets are suitable for work with microbiological agents assigned to biosafety levels 1, 2 and 3.
  • 28. class II Biological safety cabinets • 70% re circulated air; 30 % exhausted into the room • 75 fpm intake
  • 29. • Like Class I safety cabinets, Class II cabinets have a stream of inward air moving into the cabinet. This is known as the inflow and it prevents the aerosol generated during microbiological manipulations to escape through the front opening. • However, unlike Class I cabinets, the inflow on Class II cabinets flows through the front inlet grille, near the operator. None of the unfiltered inflow air enters the work zone of the cabinet, so the product inside the work zone is not contaminated by the outside
  • 30. • A feature unique to Class II cabinets is a vertical laminar (unidirectional) HEPA-filtered air stream that descends downward from the interior of the cabinet. This continuously flushes the cabinet interior of airborne contaminants and protects samples being handled within the cabinet from contamination and is known as the downflow.
  • 31. DIFFERENCEBETWEENVARIOUSCLASS2 BIOSAFETYCABINETS • The differences between the various Class II cabinets available lie primarily with the percentage of air exhausted to that of air re-circulated from the common air plenum. In addition, different Class II cabinets have different means of cabinet exhaust. Some cabinets may exhaust air directly back to the laboratory, while others may exhaust air through a dedicated ductwork system to the external environment.
  • 32. • Despite these differences, all Class II cabinets, like Class I cabinets, protect both the operator and environment from exposure to biohazards. • In addition, Class II cabinets also protect product samples from contamination during microbiological manipulations within the cabinet interior and are all suitable for work with agents assigned to biosafety levels 1, 2 and 3.
  • 33. classII typeA (A1/A2) Biologicalsafety cabinets • 70% re circulated air; 30 % exhausted into the room • 75 fpm intake Class II Type A1
  • 34. Class II Type A2 •70% re circulated air; 30 % exhausted into the room • 100 fpm intake
  • 35. • The Class II Type A biological safety cabinet is the most common Class II cabinet. It is also the most common safety cabinet of all the different types available. It has a common plenum from which 30% of air is exhausted, and 70% re- circulated to the work area as the downflow. • Type A cabinets exhaust air directly back to the laboratory, and they may contain positive pressure contaminated plenums. When toxic chemicals must be employed as an adjunct to microbiological processes, these cabinets should not be used. Exhaust HEPA filtration only removes airborne aerosols including biohazards, and not chemical fumes.
  • 36. • The Class II Type A1 has the positively-pressurized contaminated plenum bordering the ambient environment, and therefore is less safe than the Class II Type A2 that has a negative pressure surrounding the positively pressurized contaminated plenum. In case there is a leakage on the positive plenum, the leaking aerosol will be pulled by the negative pressure back to the positive plenum, and it will not leak out. Because of the safety issue, the Type A1 design is now considered obsolete.
  • 37. • In the A2 cabinet, about 70% of air from the positive plenum is recirculated as downflow, and the remaining 30% is discharged to the lab through the exhaust filter.
  • 38. 38 class II typeB1 Biological safety cabinets • Class II Type B1 • 40% re circulated air; 60 % exhausted • 100 fpm intake
  • 39. • Type B1 cabinets also have a dedicated exhaust feature that eliminates re-circulation when work is performed towards the back within the interior of the cabinet. Toxic chemicals employed as an adjunct to microbiological processes should only be employed if they do not interfere with work when re- circulated in the downflow.
  • 40. Class II BSC • Class II Type B2 • 0% re circulated air; 100 % exhausted • 100 fpm intake 40
  • 41. • In the Class II Type B2 cabinet all inflow and downflow air is exhausted after HEPA filtration to the external environment without recirculation within the cabinet. Type B2 cabinets are suitable for work with toxic chemicals employed as an adjunct to microbiological processes under all circumstances since no re-circulation occurs.
  • 42. • In theory, Type B2 cabinets may be considered to be the safest of all Class II biological safety cabinets since the total exhaust feature acts as a fail-safe in the event that the downflow and / or exhaust HEPA filtration systems cease to function normally. However, Class II Type B2 cabinets are, in practice, difficult to install, balance and maintain
  • 44. • The Class III biological safety cabinet provides an absolute level of safety, which cannot be attained with Class I and Class II cabinets. All Class III cabinets are usually of welded metal construction and are designed to be gas tight. • Work is performed through glove ports in the front of the cabinet. During routine operation, negative pressure relative to the ambient environment is maintained within the cabinet. This provides an additional fail-safe mechanism in case physical containment is compromised.
  • 45. • On all Class III cabinets, a supply of HEPA filtered air provides product protection and prevents cross contamination of samples. Exhaust air is usually HEPA filtered and incinerated. Alternatively, double HEPA filtration with two filters in series may be utilized. • Materials are transferred into the cabinet using a pass-through unit installed at the side of the work area. Class III cabinets usually exhaust air back to the laboratory; however, air may also be exhausted via a dedicated ductwork system to the external environment. When a dedicated ductwork system is employed, they are also suitable for work employing toxic chemicals as an adjunct to microbiological processes.
  • 46. • All Class III biological safety cabinets are suitable for work with microbiological agents assigned to biosafety levels 1, 2, 3 and 4. They are frequently specified for work involving the most lethal biological hazards.
  • 47. HEPA & ULPA Filter • The HEPA filter is the heart of the biological safety cabinet. The HEPA filter is a disposable dry-type filter, constructed of borosilicate microfibers cast into a thin sheet, much like a piece of paper. • • HEPA: high efficiency Particulate air: Modern “American- convention” HEPA: 99.99% at 0.3 microns • • ULPA: Ultra Low Penetration air: Modern “American- convention” ULPA: 99.999% at 0.12 microns
  • 48. • HEPA/ ULPA filters are designed to remove a broad range of airborne contaminants, including: • • Fine dust • • Smoke • • Bacteria (typical size: 500 to 0.3 microns) • • Soot • • Pollen • • Radioactive particles
  • 49. Safe use of biosafety cabinet in laboratory • 1. Know Your Airflow • Maximize potential protection by making sure you know how filtered and contaminated air, from both work surface and room, passes through your cabinet. • 2. Work at the Proper Sash Level • Your BSC will be certified for safe operation at a certain window sash height – most cabinets will indicate where the window should be placed during work activities. It is crucial to work at the certified window sash height to ensure that the balance between inflow and down flow velocities is maintained. Excessive inflow can cause contaminated air to enter the work area; excessive down flow can cause contaminated air to enter the laboratory as well as creating turbulent air in the work area, resulting in product contamination.
  • 50. • 3. Never Block the Airflow Grill • The airflow grill at the front of your cabinet performs the important job of separating the clean air inside the cabinet from the contaminated laboratory air outside. Take care to avoid covering the air grill at the front of the cabinet with equipment, and even with your arms, elbows or hands, as this can compromise airflow integrity, resulting in contaminants entering the sterile work zone inside the cabinet, or substances from inside the cabinet leaking out into the laboratory. • 4. Minimize Movement • The best practice for working in your BSC is to move your arms in and out slowly and perpendicular to the face of the opening of the cabinet to minimize the disruption of airflow within the cabinet. It is also important to organize the cabinet workspace so that minimal arm movement is required; and make sure to take care when walking near the cabinet or opening and closing doors – rapid movement near the BSC may disrupt the cabinet air barrier.
  • 51. • 5. Reduce Splatter • Use good microbiological techniques to reduce the potential risk of creating splatter or aerosol generation. Class II cabinets are designed so that horizontally nebulized spores will be captured by the downward flowing cabinet air within 14 inches of travel. As a general rule of thumb, clean materials should be kept at least 12 inches away from aerosol generating activities to prevent cross-contamination. • 6. Know Your Work Area • The middle third of the BSC work surface is the ideal area to be used for your experiments and all operations should be performed at least 4 inches from the front grill. Materials placed in the cabinet can cause disruption in airflow, resulting in turbulence, possible cross- contamination, and/or breach of containment. It is therefore important that only the materials required for immediate work are placed in the cabinet work area, and that these are placed as far back as possible, but still within reach. Bulky items such as biohazard bags should be placed to one side in the interior of the cabinet, while aerosol-generating equipment such as vortex mixers or centrifuges should be placed toward the rear of the cabinet.
  • 52. • 7. Work from Clean to Contaminated • Active work should flow from a clean to a contaminated area across the work surface to limit the movement of contaminated equipment over clean items. • 8. Working with Tubes & Bottles • Open tubes and bottles should not be held in a vertical position and their caps should not be placed on toweling. Caps should be placed back on the tube or bottle as soon as possible.
  • 53. • 9. Working with Petri Dishes • Then working with Petri dishes, the lid should be held above the open sterile surface to minimize the direct impact of downward air. • 10. Working with Aspirator Bottles or Suction Flasks • spirator bottles or suction flasks should be connected to an overflow collection flask containing appropriate disinfectant and an in-line HEPA or equivalent filter.
  • 54. • 11. Dress for the Occasion • Proper dress is also essential for your personal protection as well as your product and working environment. Laboratory coats should be worn buttoned over street clothing, protective eyewear should be on at all times and latex or nitrile gloves are necessary when handling culture, contaminated surfaces, or equipment. • 12. Make Your Setup Ergonomic • Optimize your setup to improve your safety. For example, use a good laboratory chair and sit with 90-degree seating and knee angles to reduce your risk of workplace injury. Also make sure to work with straight wrists and arrange your work area properly so you do not overextend your arms – accessories such as armrests and footrests can be used to increase comfort and support.