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Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
Laboratory Biosafety 2007
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Laboratory Biosafety 2007

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Described different bisafety levels

Described different bisafety levels

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  • In his 1979 review, Pike concluded that "the knowledge, the techniques, and the equipment to prevent most laboratory infections are available." In the United States, however, no single code of practice, standards, guidelines, or other publication provided detailed descriptions of techniques, equipment, and other considerations or recommendations for the broad scope of laboratory activities conducted with a variety of indigenous and exotic infectious agents. The booklet, Classification of Etiologic Agents on the Basis of Hazard, served as a general reference for some laboratory activities utilizing infectious agents. This booklet, and the concept of categorizing infectious agents and laboratory activities into four classes or levels, served as a basic format for earlier editions of Biosafety in Microbiological and Biomedical Laboratories (BMBL). The fourth edition of the BMBL continues to specifically describe combinations of microbiological practices, laboratory facilities, and safety equipment, and to recommend their use in four categories or biosafety levels of laboratory operation with selected agents infectious to humans.
  • Specimens shall be transported in a manner to prevent contamination of workers, patients, environment Use appropriate packing containers Follow national and international regulations
  • A number of safety references are available (CDC, WHO, ISO). More detail of what is discussed in this presentation can be found in any of these references. Details on: Safety Responsibilities Management Staff Facility Management Design Identification of hazards Housekeeping practices Biologic Safety Chemical Safety This presentation focuses primarly on biosafety
  • The term "containment" is used in describing safe methods for managing infectious materials in the laboratory environment where they are being handled or maintained. The purpose of containment is to reduce or eliminate exposure of laboratory workers, other persons, and the outside environment to potentially hazardous agents. Primary containment, the protection of personnel and the immediate laboratory environment from exposure to infectious agents, is provided by both good microbiological technique and the use of appropriate safety equipment. The use of vaccines may provide an increased level of personal protection. Secondary containment, the protection of the environment external to the laboratory from exposure to infectious materials, is provided by a combination of facility design and operational practices. Therefore, the three elements of containment include laboratory practice and technique, safety equipment, and facility design. The risk assessment of the work to be done with a specific agent will determine the appropriate combination of these elements.
  • The recommended biosafety level(s) for the organisms represent those conditions under which the agent ordinarily can be safely handled. The laboratory director is specifically and primarily responsible for assessing the risks and appropriately applying the recommended biosafety levels. When specific information is available to suggest that virulence, pathogenicity, antibiotic resistance patterns, vaccine and treatment availability, or other factors are significantly altered, more (or less) stringent practices may be specified.
  • Purposes of safety cabinets: Product protection Personal protection Environmental protection
  • Access to the laboratory is limited or restricted at the discretion of the laboratory director when experiments or work with cultures and specimens are in progress. A biohazard sign can be posted at the entrance to the laboratory whenever infectious agents are present. The sign may include the name of the agent(s) in use and the name and phone number of the investigator. Eating, drinking, smoking, handling contact lenses, applying cosmetics, and storing food for human use are not permitted in the work areas. Persons who wear contact lenses in laboratories should also wear goggles or a face shield. Food is stored outside the work area in cabinets or refrigerators designated and used for this purpose only.
  • Persons wash their hands after they handle viable materials, after removing gloves, and before leaving the laboratory. What can be done in the absence of handwashing facilities? Use antiseptic hand cleaner and clean towels.
  • Provide lockable doors for facilities that house restricted agents. Consider locating new laboratories away from public areas. The laboratory is designed so that it can be easily cleaned. Carpets and rugs in laboratories are inappropriate. Bench tops are impervious to water and are resistant to moderate heat and the organic solvents, acids, alkalis, and chemicals used to decontaminate the work surfaces and equipment. Laboratory furniture is capable of supporting anticipated loading and uses. Spaces between benches, cabinets, and equipment are accessible for cleaning. Chairs and other furniture used in laboratory work should be covered with a non-fabric material that can be easily decontaminated.
  • Properly maintained biological safety cabinets, preferably Class II, or other appropriate personal protective equipment or physical containment devices are used whenever:
  • A high degree of precaution must always be taken with any contaminated sharp items, including needles and syringes, slides, pipettes, capillary tubes, and scalpels. Needles and syringes or other sharp instruments should be restricted in the laboratory for use only when there is no alternative, such as parenteral injection, phlebotomy, or aspiration of fluids from laboratory animals and diaphragm bottles. Plasticware should be substituted for glassware whenever possible. Used disposable needles must not be bent, sheared, broken, recapped, removed from disposable syringes, or otherwise manipulated by hand before disposal; rather, they must be carefully placed in conveniently located puncture-resistant containers used for sharps disposal. Non-disposable sharps must be placed in a hard-walled container for transport to a processing area for decontamination, preferably by autoclaving.
  • Broken glassware must not be handled directly by hand, but must be removed by mechanical means such as a brush and dustpan, tongs, or forceps. Containers of contaminated needles, sharp equipment, and broken glass are decontaminated before disposal, according to any local, state, or federal regulations.
  • Laboratory equipment and work surfaces should be decontaminated with an effective disinfectant on a routine basis, after work with infectious materials is finished, and especially after overt spills, splashes, or other contamination by infectious materials. Contaminated equipment must be decontaminated according to any local, state, or federal regulations before it is sent for repair or maintenance or packaged for transport in accordance with applicable local, state, or federal regulations, before removal from the facility. Spills and accidents that result in overt exposures to infectious materials are immediately reported to the laboratory director. Medical evaluation, surveillance, and treatment are provided as appropriate and written records are maintained.
  • Laboratory equipment and work surfaces should be decontaminated with an effective disinfectant on a routine basis, after work with infectious materials is finished, and especially after overt spills, splashes, or other contamination by infectious materials. Contaminated equipment must be decontaminated according to any local, state, or federal regulations before it is sent for repair or maintenance or packaged for transport in accordance with applicable local, state, or federal regulations, before removal from the facility. Spills and accidents that result in overt exposures to infectious materials are immediately reported to the laboratory director. Medical evaluation, surveillance, and treatment are provided as appropriate and written records are maintained. Animals not involved in the work being performed are not permitted in the lab.
  • The laboratory is separated from areas that are open to unrestricted traffic flow within the building, and access to the laboratory is restricted. Passage through a series of two self-closing doors is the basic requirement for entry into the laboratory from access corridors. Doors are lockable. A clothes change room may be included in the passageway. Air flow within the laboratory is directional, single-pass and has 10-12 complete air changes per hour. Typical BSL 2 laboratories are designed with 6-8 complete air changes per hour.
  • The interior surfaces of walls, floors, and ceilings of areas where BSL-3 agents are handled are constructed for easy cleaning and decontamination. Seams, if present, must be sealed. Walls, ceilings, and floors should be smooth, impermeable to liquids and resistant to the chemicals and disinfectants normally used in the laboratory. Floors should be monolithic and slip-resistant. Consideration should be given to the use of coved floor coverings. Penetrations in floors, walls, and ceiling surfaces are sealed. Openings such as around ducts and the spaces between doors and frames are capable of being sealed to facilitate decontamination. Bench tops are impervious to water and are resistant to moderate heat and the organic solvents, acids, alkalis, and those chemicals used to decontaminate the work surfaces and equipment.
  • HEPA-filtered exhaust air from a Class II biological safety cabinet can be recirculated into the laboratory if the cabinet is tested and certified at least annually. When exhaust air from Class II safety cabinets is to be discharged to the outside through the building exhaust air system, the cabinets must be connected in a manner that avoids any interference with the air balance of the cabinets or the building exhaust system (e.g., an air gap between the cabinet exhaust and the exhaust duct). When Class III biological safety cabinets are used they should be directly connected to the exhaust system. If the Class III cabinets are connected to the supply system, it is done in a manner that prevents positive pressurization of the cabinets.
  • All manipulations of infectious materials, necropsy of infected animals, harvesting of tissues or fluids from infected animals or embryonate eggs , etc., are conducted in a Class II or Class III biological safety cabinet . When a procedure or process cannot be conducted within a biological safety cabinet, then appropriate combinations of personal protective equipment (e.g., respirators, face shields) and physical containment devices (e.g., centrifuge safety cups or sealed rotors) are used.
  • All manipulations of infectious materials, necropsy of infected animals, harvesting of tissues or fluids from infected animals or embryonate eggs , etc., are conducted in a Class II or Class III biological safety cabinet. When a procedure or process cannot be conducted within a biological safety cabinet, then appropriate combinations of personal protective equipment (e.g., respirators, face shields) and physical containment devices (e.g., centrifuge safety cups or sealed rotors) are used.
  • Housekeeping procedures may include: details for preparing disinfectants: frequency of cleaning work stations. Ex: Use O.1% chlorine-based disinfectant to clean bench after completion of work for the morning and then at the end of the day Housekeeping also includes laundry procedures for labcoats.
  • Washing Hands   Always wash hands with soap and water after handling specimens and containers.
  • Transcript

    • 1. An overview of Laboratory Safety Biosafety Dr. Ashok Rattan, Medical Microbiologist & Laboratory Director, CAREC
    • 2. Before entering the water to swim you must learn all you can about the sharks that may lurk there SAFETY IN THE MYCOBACTERIOLOGY LABORATORY
    • 3. <ul><li>Protection: </li></ul><ul><ul><li>workers </li></ul></ul><ul><ul><li>“ products” </li></ul></ul><ul><ul><li>co-workers </li></ul></ul><ul><ul><li>lab support personnel </li></ul></ul><ul><ul><li>environment </li></ul></ul>Introduction Why Biosafety Practices? 2.1
    • 4. Safety practices should be applied throughout the testing process: <ul><li>Pre- analytical </li></ul><ul><ul><li>Specimen collection </li></ul></ul><ul><ul><li>Specimen preparation </li></ul></ul><ul><ul><li>Specimen transport </li></ul></ul><ul><li>Analytical </li></ul><ul><ul><li>Testing </li></ul></ul><ul><li>Post-analytical </li></ul><ul><ul><li>Disposal </li></ul></ul>
    • 5. Examples of Laboratory Hazards <ul><li>Growth of microorganisms </li></ul><ul><li>Water baths </li></ul><ul><li>Aerosols, splashing, tube breakage </li></ul><ul><li>Centrifuge </li></ul><ul><li>Accidental inoculation, aerosol, spillage </li></ul><ul><li>Needles </li></ul>Hazard Operation / Equipment
    • 6. Safety Resources
    • 7. Safety Resources
    • 8. Introduction Chain of Infection Reservoir of pathogen Portal of escape Transmission Route of entry/infectious dose Susceptible host Incubation period Risk Assessment PPE Immunization Surveillance Practices/ Equipment
    • 9. <ul><li>BSL1 - agents not known to cause disease. </li></ul><ul><li>BSL2 - agents associated with human disease. </li></ul><ul><li>BSL3 - indigenous/exotic agents with potential for aerosol transmission; disease may have serious or lethal consequences. </li></ul><ul><li>BSL4 - dangerous/exotic agents which pose high risk of life-threatening disease. </li></ul>Principles Biosafety Levels of agents 2.1
    • 10. Designing for Safety <ul><li>General requirements </li></ul><ul><ul><li>Facility design </li></ul></ul><ul><ul><li>Water supply/sinks for hand washing </li></ul></ul><ul><ul><li>Ventilation </li></ul></ul><ul><ul><li>Standard lab practices </li></ul></ul><ul><li>Safety equipment </li></ul><ul><ul><li>Personal protective equipment (PPE) </li></ul></ul><ul><ul><li>Biosafety cabinets </li></ul></ul>
    • 11. Biosafety Level 1 Standard Microbiological Practices 1/3 <ul><li>Restrict or limit access when working </li></ul><ul><li>Prohibit eating, drinking and smoking in the laboratory </li></ul><ul><li>Pipetting by mouth strictly forbidden </li></ul>2.3
    • 12. Biosafety Level 1 Standard Microbiological Practices 2/3 2.3
    • 13. Standard practices also include: 3/3 <ul><li>Keep work areas uncluttered and clean </li></ul><ul><li>No food in lab refrigerator </li></ul><ul><li>Minimize splashes and aerosols </li></ul><ul><li>Decontaminate work surfaces daily </li></ul><ul><li>Maintain insect & rodent control program </li></ul>
    • 14. Biosafety Level 2 Facility Design (Secondary Barriers) 1/8 <ul><li>Requirements: </li></ul><ul><ul><li>Laboratories have lockable doors </li></ul></ul><ul><ul><li>Sink for hand washing </li></ul></ul><ul><ul><li>Work surfaces easily cleaned </li></ul></ul><ul><ul><li>Bench tops are impervious to water </li></ul></ul><ul><ul><li>Sturdy furniture </li></ul></ul>2.4
    • 15. Biosafety Level 2 Laboratory Facilities (Secondary Barriers) 2/8 <ul><li>BSL-1 Facilities PLUS: </li></ul><ul><ul><li>Autoclave available </li></ul></ul><ul><ul><li>Eyewash station available </li></ul></ul>2.4
    • 16. <ul><li>Requirements: </li></ul><ul><ul><li>Location - separated from public areas </li></ul></ul><ul><ul><li>Structure - normal construction </li></ul></ul><ul><ul><li>Ventilation - directional </li></ul></ul>Biosafety Level 2 Facility Construction (Secondary Barrier) 3/8 2.4
    • 17. Biosafety Level 2 Safety Equipment (Primary Barriers) 4/8 <ul><li>In addition to BSL-1: </li></ul><ul><li>Use biosafety cabinets (class II) for work with infectious agents involving: </li></ul><ul><ul><li>Aerosols and splashes </li></ul></ul><ul><ul><li>Large volumes </li></ul></ul><ul><ul><li>High concentrations </li></ul></ul>2.4
    • 18. Biosafety Level 2 Special Practices 5/8 <ul><li>Needles & Sharps Precautions </li></ul><ul><ul><li>Use sharps containers </li></ul></ul><ul><ul><li>DON’T break, bend, re-sheath or reuse </li></ul></ul><ul><li>syringes or needles </li></ul>2.4
    • 19. Biosafety Level 2 Special Practices 6/8 <ul><li>Needles & Sharps Precautions </li></ul><ul><li>DON’T place needles or sharps in office waste containers </li></ul>2.4
    • 20. Biosafety Level 2 Special Practices 7/8 <ul><li>Needles and Sharps Precautions </li></ul><ul><ul><li>DON’T t ouch broken glass with hands </li></ul></ul>2.4
    • 21. <ul><li>Identify “clean” and “contaminated” areas </li></ul><ul><ul><li>Use appropriate warning signs </li></ul></ul><ul><li>Decontaminate work surfaces </li></ul><ul><li>Report spills and accidents </li></ul><ul><li>Remove gloves, lab coats before leaving work area </li></ul><ul><li>No animals in laboratories </li></ul>Biosafety Level 2 Special Practices 8/8 2.4
    • 22. Biosafety Level 3 Laboratory Facilities (Secondary Barriers) 1/5 <ul><li>BSL-1 and 2 Facilities PLUS: </li></ul><ul><ul><li>Separate building or isolated zone </li></ul></ul><ul><ul><li>Double door entry </li></ul></ul><ul><ul><li>Directional inward airflow </li></ul></ul><ul><ul><li>Single-pass air; 10-12 air changes/hour </li></ul></ul>2.5
    • 23. Biosafety Level 3 Laboratory Facilities (Secondary Barriers) 2/5 <ul><li>BSL-1 and 2 Facilities PLUS (cont.): </li></ul><ul><ul><li>Enclosures for aerosol generating equipment </li></ul></ul><ul><ul><li>Room penetrations sealed </li></ul></ul><ul><ul><li>Walls, floors and ceilings are water resistant for easy cleaning </li></ul></ul>2.5
    • 24. Biosafety Level 3 Safety Equipment (Primary Barriers) 3/5 <ul><li>BSL-1 and 2 Safety Equipment PLUS: </li></ul><ul><ul><li>BSC class II or III to manipulate infectious material </li></ul></ul>2.5
    • 25. Biosafety Level 3 Safety Equipment (Primary Barriers) 4/5 <ul><li>BSL-1 and 2 Safety Equipment PLUS: </li></ul><ul><ul><li>Respiratory protection may be indicated </li></ul></ul>2.5
    • 26. <ul><li>BSL-2 Special Practices PLUS: </li></ul><ul><ul><li>Work in certified BSC </li></ul></ul><ul><ul><li>Use bioaerosol- containing equipment </li></ul></ul><ul><ul><li>Decontaminate spills promptly </li></ul></ul>Biosafety Level 3 Special Practices 5/5 2.5
    • 27. Biosafety Level 4 – Maximum Containment <ul><li>BSL -3 practices plus: </li></ul><ul><ul><li>Clothing change before entering laboratory </li></ul></ul><ul><ul><li>Shower on exit </li></ul></ul><ul><ul><li>All materials decontaminated on exit from facility </li></ul></ul><ul><li>Safety Equipment: </li></ul><ul><ul><li>Class III Biosafety cabinet </li></ul></ul><ul><ul><li>Class I or II biosafety cabinet </li></ul></ul><ul><ul><li>WITH full-body, air supplied, </li></ul></ul><ul><ul><ul><li>positive personnel suit </li></ul></ul></ul>
    • 28. Biological Waste <ul><li>Types </li></ul><ul><ul><li>cultures, stocks, isolates </li></ul></ul><ul><ul><li>materials containing or contaminated with blood </li></ul></ul><ul><ul><li>sharps </li></ul></ul><ul><ul><li>pipettes, wrappers, tips </li></ul></ul><ul><ul><li>All materials used in the lab </li></ul></ul>
    • 29. Specimen Disposal
    • 30. Decontamination <ul><li>Sterilization </li></ul><ul><li>Disinfection </li></ul>
    • 31. Decontamination Definition <ul><li>Sterilization </li></ul><ul><ul><li>The use of a physical or chemical procedure to destroy all microbial life, including large numbers of highly resistant bacterial spores. </li></ul></ul>
    • 32. <ul><li>Disinfection </li></ul><ul><ul><li>The use of a physical or chemical procedure to virtually eliminate all recognized pathogenic microorganisms but not all microbial forms (bacterial endospores) on inanimate objects. </li></ul></ul>Decontamination Definition
    • 33. Decontamination Methods <ul><li>Heat </li></ul><ul><li>Chemical </li></ul><ul><li>Radiation </li></ul>
    • 34. <ul><li>Types </li></ul><ul><ul><li>Moist – steam </li></ul></ul><ul><ul><li>Dry </li></ul></ul><ul><ul><li>Incineration </li></ul></ul><ul><li>*The most effective method of sterilization </li></ul>Decontamination Heat
    • 35. <ul><li>Types </li></ul><ul><ul><li>Liquids, i.e. chlorox, hydrogen peroxide </li></ul></ul><ul><ul><li>Gases, i.e. ethylene oxide </li></ul></ul>Decontamination Chemical
    • 36. <ul><li>General Lab Use - Hypochlorite Solutions </li></ul><ul><ul><li>Large Spills/Large Organic Load </li></ul></ul><ul><ul><ul><li>undiluted from bottle </li></ul></ul></ul><ul><ul><li>Small Spills/Virus Inactivation </li></ul></ul><ul><ul><ul><li>10% - 1:9 </li></ul></ul></ul><ul><ul><li>General Surface Disinfection </li></ul></ul><ul><ul><ul><li>1% - 1:99 </li></ul></ul></ul>Decontamination Chemical
    • 37. In case of a spill <ul><li>Wear disposable gloves </li></ul><ul><li>Cover large blood spill with paper towels and soak with 1% (10000 ppm) of household bleach and allow to stand for at least 5 minutes </li></ul><ul><li>Small spill - wipe with paper towel soaked in 1% bleach </li></ul><ul><li>Discard contaminated towels in infective waste containers </li></ul><ul><li>Wipe down the area with clean towels soaked in a same dilution of household bleach </li></ul>
    • 38. Safety Documentation & Records <ul><li>Laboratory Safety Manual - Policies and Procedures </li></ul><ul><li>Sample Contents: </li></ul><ul><ul><li>Housekeeping </li></ul></ul><ul><ul><li>Personal protection </li></ul></ul><ul><ul><li>Safe decontamination of equipment </li></ul></ul><ul><ul><li>Decontamination & Waste Disposal </li></ul></ul><ul><ul><li>Emergency procedures </li></ul></ul><ul><ul><ul><ul><li>In-lab first aid </li></ul></ul></ul></ul><ul><ul><ul><ul><li>Accidental injury </li></ul></ul></ul></ul><ul><ul><ul><ul><li>Post exposure prophylaxis </li></ul></ul></ul></ul><ul><ul><ul><ul><li>Contacts </li></ul></ul></ul></ul><ul><ul><li>Personnel responsibilities </li></ul></ul><ul><ul><ul><li>Hygiene </li></ul></ul></ul>
    • 39. Action Plan for Implementing Safety Practices <ul><li>Identify hazards </li></ul><ul><li>Assess level of risks </li></ul><ul><ul><li>Prioritize risk </li></ul></ul><ul><li>Establish and implement safety polices and procedures </li></ul><ul><li>Conduct safety specific training </li></ul><ul><ul><li>Must be a priority </li></ul></ul><ul><ul><li>Communication is key </li></ul></ul><ul><li>Perform regular audits and assessments </li></ul>
    • 40. In Case of Exposure <ul><li>Be ready for the emergency before hand </li></ul><ul><li>- Familiar with exposure specific policies </li></ul><ul><li>- Conduct drills </li></ul><ul><li>- Keep post exposure medicines available </li></ul><ul><li>- Check periodically for stock and expiry of medicines </li></ul><ul><li>Report immediately </li></ul><ul><li>Go to the nearest, first available doctor </li></ul>
    • 41. Post Exposure <ul><li>Write a report and reasons for accident </li></ul><ul><li>Actions taken to avoid future accidents </li></ul><ul><li>Training </li></ul>
    • 42. SAFETY IN THE MYCOBACTERIOLOGY LABORATORY
    • 43.  

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