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Vivek bhat TMC vivek bhat tmc. Potential Laboratory Hazards Chemicals Includes different classes of hazardous chemicals Biohazards Cells, animals, biological.

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Presentation on theme: "Vivek bhat TMC vivek bhat tmc. Potential Laboratory Hazards Chemicals Includes different classes of hazardous chemicals Biohazards Cells, animals, biological."— Presentation transcript:

1 vivek bhat TMC vivek bhat tmc

2 Potential Laboratory Hazards Chemicals Includes different classes of hazardous chemicals Biohazards Cells, animals, biological / patient samples, viruses, bacteria Allergens Chemical, animal, latex Radioactive Material Physical / Equipment Hazards Electrical, sharps, hi/low temperature and pressure Mixed hazards & Multiple simultaneous hazards 2 vivek bhat tmc

3 Biohazard: Any agent of biological origin that has the capacity to produce harmful effects in humans. Biosafety: Safety against biohazards A laboratory acquired infection is defined as one that resulted from laboratory work, whether it occurred in a laboratory worker or in another person who happened to be exposed as a result of research or clinical work with infectious agents. Antiseptic :– A substance that inhibits the growth and development of micro-organisms without necessarily killing them. Antiseptics are usually applied to body surfaces. Disinfectant :– A chemical or mixture of chemicals used to kill microorganisms, but not necessarily spores. Disinfectants are usually applied to inanimate surfaces or objects. 3 vivek bhat tmc

4 Biocide – A general term for any agent that kills organisms. Sporocide – A chemical or mixture of chemicals used to kill microorganisms and spores. Decontamination – Any process for removing and/or killing microorganisms. The same term is also used for removing or neutralizing hazardous chemicals and radioactive materials. Disinfection – A physical or chemical means of killing microorganisms, but not necessarily spores. Sterilization – A process that kills and/or removes all classes of microorganisms and spores. 4 vivek bhat tmc

5 Any agent of biological origin that has the capacity to produce harmful effects in humans. Examples of biohazards: Micro-organsims such as bacteria, viruses, fungi and parasites and their toxins. Blood and body fluids as well as tissues from humans and animals. Transformed cell lines and certain types of nucleic acids. Every infectious microbial agent which has been studied in the laboratory has caused infection in lab personnel Fewer than 20% of all LAIs were associated with a known accident. 80% go unknown or unrecognized 5 vivek bhat tmc

6 Q fever Brucellosis Typhoid fever Hepatitis Tularemia Tuberculosis Dermatomycosis Venezuelan equine encephalitis Typhus Psittacosis Coccidioidomycosis. Leptospirosis C. difficile E. coli O157:H7 N. meningitidis Salmonella Shigella HIV HBV HCV 6 vivek bhat tmc

7 RouteLab Practices and/or Accidents Protection Inhalation Procedure s that produce aerosols vapours, dust, mists, gases and biological agents fume hood, masks, respirator or BSC as appropriate Ingestion Mouth pipetting Splashes into mouth Contaminated hands /utensils Eating, drinking, smoking etc Leaking contaminated items NO eating or drinking or makeup application in the lab. NO mouth pipetting Inoculation Needle stick injury Cuts from sharps Animal and insect bites limit use of sharps and if possible handle indirectly with forceps. Use sharps container Other - contact Spills and splashes Contact with contaminated items Transfer by hand –to-face actions Standard Precautions and GMP 7 vivek bhat tmc

8 WHO recommends that the Health Authorities of every country make lists of organisms in the different risk groups, relevant to the local circumstances, so that appropriate precautions may be applied. Risk group 1 : Organisms in this group represents low risk to the lab worker & to members of the community. They are unlikely to cause human disease. (e.g. saprophytic bacteria, animal/plant organisms) Risk group 2 : Organisms in this group offer a moderate risk to the lab worker limited risk to members of the community. 8 vivek bhat tmc

9 Risk group 2 ….cont They can cause human disease but preventive measures & treatment /vaccines etc are available risk of spread in the community is not great E.g : staph, strep., enterobacteriaceae, clostridia, vibrio, polio, coxsackie & hepatitis viruses. Risk group 3 : Presents high risk to Laboratory worker Low risk to community Vaccines & treatment available for most pathogens Eg ; Brucella, Mycobacterium tb, S. typhi, Pasteurella, Francisella, many arboviruses, Ricketttsiae, Chlamydia, HIV. 9 vivek bhat tmc

10 Risk group 4 : High risk to Lab worker & community. Cause serious disease & spreads easily & rapidly in the community No vaccines or chemotherapy agents available. E.g : Hemorrhagic fever viruses including Marburg, Lassa, Ebola, & some encephalitis & arboviruses. 10 vivek bhat tmc

11 Basic Laboratory (Bio-safety level 1) Standard Good Laboratory practices (GLP) required Simplest kind – BSCs not required. Basic Laboratory (Bio-safety level 2) Suitable for Risk group 2 GLP as outlined with all the standard precautions Training Lab personnel in handling infectious organisms & suspensions. BSCs ( Class 1 or Class 2) in addition to all other equipment including autoclaves Employee health policy (GLP includes aseptic tech, personal hygiene area clean- liness, following standard precautions and PPE, etc) 11 vivek bhat tmc

12 Containment Laboratory (Biosafety Level 3) Suitable for Risk group 3 Controlled access by authorized staff only Special Lab design & engineering & HVAC with careful control of air movement. Wearing of special protective clothing & accessories. Laminar flows, Biological safety cabinets - class 2 Effective Employee Health policy with baseline sera stored for comparison with acute sera. 12 vivek bhat tmc

13 Containment Laboratory (Biosafety Level 4) Suitable for Risk group 4 Includes all facilities of Level 3 plus Separate buildings/units with strict controlled access through air locks & exit decontamination showers & pressure gradient rooms. Special training for staff. Requirement of BSC Class 3 Effective Employee Health Policy with baseline sera stored for comparison with acute sera. Most are Research/ Defense Laboratories 13 vivek bhat tmc

14 BiologicalSafetyCabinet SterilGARD® III Advance, The Baker Company 14 vivek bhat tmc

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21 The use and limitations of biological safety cabinets should be explained to all potential users. Written protocols or safety or operations manuals should be issued to staff. In particular, it must be made clear that the cabinet will not protect the operator from spillage, breakage or poor technique. Do not use the cabinet unless it is working properly. Do not open the glass viewing panel when the cabinet is in use. Apparatus and materials in the cabinet must be kept to a minimum. Air circulation at the rear plenum must not be blocked. 21 vivek bhat tmc

22 Bunsen burners must not be used in the cabinet. The heat produced will distort the airflow and may damage the filters. An electric micro- incinerator is permissible but sterile disposable transfer loops are better. All work must be carried out in the middle or rear part of the working surface and be visible through the viewing panel. Traffic behind the operator should be minimized. Air grills must not be blocked with notes, pipettes or other materials, as this will disrupt the airflow causing potential contamination of the material and exposure of the operator. Wipe the surface of the BSC using an appropriate disinfectant after work is completed and at the end of the day. The cabinet fan should be run for at least 5 min before beginning work and after completion of work in the cabinet. Do not place paperwork inside BSCs. 22 vivek bhat tmc

23 Use of Biohazard and safety symbols Protective clothing : Overalls, lab coats to be worn over normal clothing to protect from splashes, droplets containing micro- organisms/chemicals. Waterproof aprons may be worn when indicated, poly- cottons & flammable – resistant fabric is ideal. Laundering regularly, soiled clothing is placed in a special bag & disinfected with 1% v/v Na Hypo- chlorite & then sent to laundry. (or sent and processed as per CSSD protocol) Gloves must be worn when handling specimens of blood or body fluids or cultures & while washing lab ware. 23 vivek bhat tmc

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26 Wear gloves when you anticipate being in contact with blood or body fluids. Wear protective eyewear, masks, and gowns in addition to gloves when you think splashing of blood or body fluids could occur. 26vivek bhat tmc

27 Wear gloves when there is reasonable likelihood of hand contact with blood or other potentially infectious materials, mucous membranes or non-intact skin; when performing vascular access procedures; and when handling contaminated items or surfaces. Change disposable gloves when they become contaminated, torn or punctured. Wash hands before donning AND after removing gloves. 27 vivek bhat tmc

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29 Z87 Type Goggles O 29 vivek bhat tmc

30 Vinyl synthetic examination gloves: tasks that involve minimal stress on the glove. low risk of exposure to blood and other potentially infectious materials. Consideration needs to be given to manipulation and other stresses placed on the glove material. Suggested use: For changing bed linens. for briefly suctioning endo-tracheal secretions; for emptying emesis basins; For discontinuing an IV line; and for handling and preparing food. Not recommended: When there is moderate to high risk of exposure to blood or body fluids; for preparing, handling or administering chemotherapeutic agents. for handling chemicals or other caustic agents; for performing environmental services or housekeeping duties; If sensitivity or clinical reactivity to vinyl compounds. 30 vivek bhat tmc

31 Natural rubber latex examination gloves the gold standard in barrier protection. Natural rubber provides dexterity, tactile sensitivity, flexibility and durability. preferred for procedures and tasks considered moderate to high risk for exposure to blood and other potentially infectious materials and when a non-sterile hand covering is indicated. Latex exam gloves may need to be changed every 15 to 30 minutes depending on the task or procedure, the amount of blood and fluid exposure and the contact with needles and other sharp instruments. Suggested use: For direct patient care involving exposure to blood or other potentially infectious materials and for contact with blood and body fluid specimens or items contaminated with blood or body fluids. Not recommended: For individuals with a known or suspected allergy or clinical reactivity to natural rubber latex protein and for prolonged contact with high-level disinfectants, such as gluteraldehyde. 31 vivek bhat tmc

32 Nitrile examination gloves: Preferred choice for those individuals who have a sensitivity or clinical reactivity to latex AND may be at moderate to high risk of exposure to blood and other potentially infectious materials. Nitrile gloves typically have better chemical resistance than natural rubber latex, especially to hydrocarbon-based products (e.g., products containing mineral oil, petrolatum or lanolin). Suggested use: For personnel who are allergic or sensitive to latex and who perform tasks or procedures. involving prolonged exposure to blood, body fluids, chemo- therapeutic agents, cleaning solutions and other chemicals. Not recommended: For individuals who have Sensitivity or clinical reactivity to nitrile compounds. 32 vivek bhat tmc

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35 l) Personal health & safety measures ….cont Covering any cuts, sores, wounds etc with water proof dressing Do Not wear lab coats (and gloves) in the following areas: All offices, bathrooms, elevators, public hallway Coffee/ lunch rooms, departmental libraries Student carrel area outside of the lab Other non-lab areas of the building. No licking gum labels, putting pen/pencil in mouth/hair. Remove your gloves before using instruments, telephone, and leaving the laboratory Always wash your hands before you leave and especially before eating. 35 vivek bhat tmc

36 Dont apply cosmetics in lab Avoid wearing jewellary. (pendants, necklaces, bracelets Dont touch your face, mouth or eyes without washing hands Not eating, drinking, chewing gum, smoking, applying cosmetics or sitting on lab benches. No storing food/drink in lab refrigerators. 36vivek bhat tmc

37 Hand Hygiene Techniques 1. Alcohol hand rub: 20 to 30 sec. ( % ethanol or isopropyl alcohol) 2. Rapid decontamination of hand: no visible soiling 15 sec. (chlorhexidine 1% in alcohol) 3. Hand wash procedure (before Aseptic procedures): 1 minute (chlorhexidine - 2% handwash) 4. Surgical wash: 3-5 minutes (chlorhexidine - 4% handwash) 37 vivek bhat tmc

38 Areas Most Frequently Missed HAHS © 1999 Distribution of areas missed during hand washing 38 vive bhat tmc

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41 Do staff wash their hands enough ? No Wards16 – 48% NICU29% ICU 30 – 81% Why Dont Staff Wash their Hands (Compliance estimated at less than 50%) Skin irritation Inaccessible hand washing facilities Wearing gloves Too busy Lack of appropriate staff Being a physician (Improving Compliance with Hand Hygiene in Hospitals Didier Pittet. Infection Control and Hospital Epidemiology. Vol. 21 No. 6 Page 381) 41 vivek bhat tmc

42 Aerosolization: An aerosol ( 13 µ) is a suspension of fine solid particles or liquid droplets in a gas Small droplets evaporate & leave behind droplet nuclei consisting of bacteria or viruses which are too light to settle & they move around by air currents & ventilating systems. Particles >10 µm diameter are filtered in nose, smaller ones especially < 10 µm are inhaled right into the lungs where they initiate infection 42 vivek bhat tmc

43 Aerosolization results from Pouring supernatant fluids (from a height into a discarding container). Vigorous tapping of the tube to re-suspend a sediment. Opening cultures & rapid snap-closing of specimens or cultures. Heating a contaminated wire loop in an open burner flame. Using a long springy loop. Rapid rinsing of Pasteur pipettes. Open centrifugation, opening lid before rotation stops, opening centrifuge immediately following the breakage of tube before aerosols settle. Mouth pipetting & blowing out fluids (especially last drop) 43 vivek bhat tmc

44 Pipettes Raymond A. Lamb, LLC VWR International Beckman Coulter, Inc. Aerosol Producers Vortex Tubes 44 vivek bhat tmc

45 Prevent aerosolization Pouring infectious material safely Fluids should be poured carefully down the side of a funnel emptying in disinfectant in a jar. Opening cultures & ampoules safely Liquid films containing microorganisms form between the rims & bottoms of Petri dishes & rims & stoppers of bottles & tubes. they must be separated carefully preferably in a safety cabinet. Inoculating loops & safe looping out Loops must be fully closed - of smaller diameter - length of wire must be short (< 6mm) Use hooded burners or safety cabinets Cool the loops before use. 45 vivek bhat tmc

46 Shaking & homogenizing safely Capping the culture containers tightly while shaking, vortex mixer may be used. Avoiding infection from spillages & breakages Specimens, culture tubes & bottles should always be placed in racks so that they cannot fall over & spill their contents. Spilled material & broken culture vessels should be covered with a cloth soaked in disinfectant, left for 30 min & then cleaned up using a metal dust pan & stiff card board 46 vivek bhat tmc

47 Safe pipetting & dispensing Mouth pipetting has been the cause of many infections in lab personnel, some of which have been fatal. Serious chemical injuries may occur with chemicals. WHO recommends that mouth pipetting be banned. Low cost pipette fillers are available commercially. Air should never be blown through a liquid containing infectious agents. Infectious materials should not be mixed by alternate suction and expulsion 47 vivek bhat tmc

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49 How do Needlestick Injuries Occur? Disposing of needles Overfilling container Emptying sharps container rather than disposing once filled Administering injections Drawing blood Recapping needles. Handling trash and dirty linens. Improper disposal in regular garbage 49 vivek bhat tmc

50 Wash the area thoroughly with 2% chlorhexidine antiseptic soap solution and water. Skin sites and wounds that have been in contact with blood/body fluids shall also be immediately washed as above. Do wash mucosal surfaces like mouth or conjunctiva with warm water or saline. Do not scrub the affected area. Do not suck the area. Do not apply any caustic agents (hypochlorite etc); Do not inject antiseptic/disinfectant into the wound. Do not apply tight bandages. Baseline titres for HIV, HBsAg, HCV, Anti- HBs done depending on vaccination history. ART, HB- Ig, HBV vaccine etc provided on case to case basis. 50 vivek bhat tmc

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65 Exposure to Human Blood or Body Fluids If you are exposed to human blood or body fluids through a cut, stab or splash: – First Aid - wash immediately with soap and water (5 min) – Eyes flush with water for 10 min – Cuts, stab scrub area to bleed – Remove gloves or clothing if required Report accident and seek medical aid and follow-up 65 vivek bhat tmc

66 Spills outside of a containment device The spill is not inside of a Biological Safety Cabinet (BSC), Centrifuge, Refrigerator, Incubator, Freezer, Lab instrument etc. 1. Close off spill area to traffic, and notify coworkers. 2. If the spill may involve an aerosol, (e.g. event involving dropping material onto floor, high mechanical force, a forceful expulsion of liquid) leave the room for 30 minutes to allow aerosols to settle. 3. Remove contaminated lab coat or clothing and wash exposed skin. 4. Put on clean gloves and lab coat. 5. Prepare enough volume of a 1:10 dilution (to give 5g/l available chlorine) of chlorine bleach or other approved disinfectant to saturate the contaminated area. If dilution is not possible, undiluted household bleach can be used. However eye protection must be worn. 66 vivek bhat tmc

67 1. Contain the spill with paper towels or other absorbent material 2. Flood the spill area with disinfectant. Leave on for min. 3. Push the absorbent material at the edge of the spill into the spill's center. Add more paper towels as needed. If glass is present, do not use bare hands! Use tongs (large pieces) forceps (small pieces) followed by a dustpan to remove pieces. 4. Discard the paper towels into a regulated medical waste container. If contact with bleach occurs with skin, mucous membranes or eyes, flush area with copious amounts of water. 5. Discard gloves into regulated medical waste container. Wash hands thoroughly. Autoclave an overtly contaminated lab coat. 6. Report incident to supervisor. 67 vivek bhat tmc

68 A general all-purpose laboratory disinfectant should have a concentration of 1 g/l available chlorine. A stronger solution, containing 5 g/l available chlorine, is recommended for dealing with bio-hazardous spillage and in the presence of large amounts of organic matter. Sodium hypochlorite solutions, as domestic bleach, contain 50 g/l available chlorine (5% w/v) and should therefore be diluted 1:50 or 1:10 to obtain final concentrations of 1 g/l and 5 g/l, respectively. (tap water for dil- freshly prep soln). 1: 5 dilution ( 1% hypochlorite) may also be used for dirty conditions ( as we do in TMC) Industrial solutions of bleach have a sodium hypochlorite concentration of nearly 120 g/l and must be diluted accordingly to obtain the levels indicated above. General lab disinfectant 68 vivek bhat tmc

69 Gluteralde-- hyde 2.0 – 3.2% High/ CS Hydrogen peroxide 3.0 – 25% High/ CS ± ± Chlorine 1000p pm Cl- High ± Isopropyl alcohol 60 – 95% Int ± + -- ± + + Phenolic compounds % Int ± Iodophors 50ppm free I Int ± -- ± Q.A.Cs 0.4 – 1.6% Low ± + ± Lipophilic virusesBacterial spores Small hydrophilic viruses Bacteria Fungi Corrosive effect Shelf life > 1 week M. tuberculosis Residue Skin /Eye irritation Disinfection level GERMICIDE Dilutions 69 vivek bhat tmc

70 CBWTF- landfill after stabilization Or Recycling Mercury Spill cleanup 70 vivek bhat tmc

71 Spills in a centrifuge. Biohazardous spills in centrifuges can be quite difficult to disinfect. Some but not all centrifuges have closed rotors, buckets or other carriers with leak proof lids, designed to contain spills and allow efficient, safe emptying and decontamination. However, not all centrifuges are equipped with these containment devices. If unusual sounds from a centrifuge suggest that breakage and a spill has occurred, or, if breakage and a spill is discovered after the machine has stopped, wait at least 30 minutes after centrifuge has stopped before opening. This will allow hazardous aerosols to settle in the centrifuge. Don lab coat, gloves, and face shield prior to opening centrifuge and then open carefully to assess the situation. Use of a respirator is recommended and double gloving is advisable if glass tubes were used and broken. Attempt to determine if the spill is contained in a closed cup, bucket or tray carrier, or within a closed rotor. 71 vivek bhat tmc

72 Remove rotors and buckets to nearest biological safety cabinet for clean-up. Carefully retrieve unbroken tubes, wipe outside with disinfectant, The broken glass tube (s) must be removed with a forceps and the pieces can then be disposed of in a sharps container. After proper decontamination with disinfectant, carriers, rotors etc. can be washed with a mild detergent according to the manufacturers instructions. Thoroughly wipe the inside of the centrifuge chamber with disinfectant saturated paper towels. Allow for adequate contact time before wiping up excess liquid chlorhexidine + cetrimide (Savlon) or 70% alcohol may be used if bleach is corrosive 72 vivek bhat tmc

73 The Science: The only documented risk of transmission of infections from waste to healthcare workers is through sharps There is however a potential for transmission of several microbial infections due to dumping of untreated wastes by healthcare facilities. Mixing of a small quantity of infectious waste with municipal garbage converts the entire waste to infectious Segregation of wastes at source followed by appropriate treatment is the key to the success of a waste management strategy The Reality: Ignorance Commercialization of science Apathy The concerns: Occupational Public health Environmental 73 vivek bhat tmc

74 Hospital waste Hazardous Non-hazardous NoninfectiousInfectious KitchenRecyclables Non-sharps Patient contaminated waste Anatomical Equipment Specimens Laboratory waste Non-plastics Sharps: needles, scalpel blades, scalp veins, glass contaminated with blood Radioactive Cytotoxic drugs Toxic Chemicals Plastics PVC, PE PET, PS contaminated cotton waste, gauze, linen 74 vivek bhat tmc

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76 Category of waste Waste categoryTreatment & DisposalColor coding 1Human anatomical waste ( human organs, tissue, body parts) Incineration/deep burial yellow 2Animal waste ( animal parts, organs, body parts, bleeding parts, fluid, blood, animals used in research, waste from veterinary hospitals, colleges, animal houses) Incineration/deep burial yellow 3Microbiology and Biotechnology waste Waste from laboratory cultures, stocks, or specimens with microorganisms, live or attenuated vaccines, human & animal cell culture used in research, infectious agents from research and industrial laboratories, wastes from production of biologicals, toxins, dishes and devices used for transfer of cultures) Autoclaving/ microwaving incineration. Yellow/red 4Waste sharps: (needles, syringes, scalpels, blades, glass, etc. that may cause puncture and cuts. This includes both used and unused sharps) Disinfection/chemical treatment/autoclaving /microwaving and mutilation/ shredding Blue/white puncture proof container 76 vivek bhat tmc

77 5Discarded medicines and cytotoxic drugs: wastes comprising of outdated contaminated and discarded medicines Incineration/destruction and disposal in secured landfill Black/yellow 6Solid waste: (Items contaminated with blood and body fluids incluidng cotton dressings, soiled plaster casts, lines beddings, other materials contaminated with blood Incineration/autoclaving/ microwaving Yellow/red 7Solid Waste (wastes generated from disposable items other than the waste sharps such as tubings, catheters, IV sets etc) Disinfection by chemical treatment/autoclaving/mi crowaving and mutilation shredding Red/ blue or white bag 8Liquid Waste (waste generated from laboratory and washing, cleaning, housekeeping activities Disinfection by chemical treatment and discharge into drains -- 9Incineration ash: (ash from incineration of any biomedical waste) Disposal in municipal landfill Black 10Chemical Waste (chemicals used in production of biologicals, chemicals used in disinfection, as insecticides etc) Chemical treatment and discharge into drains for liquids and secured landfill for solids Black 77 vivek bhat tmc

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