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2012 Annual Biosafety & Chemical Safety Training.

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Presentation on theme: "2012 Annual Biosafety & Chemical Safety Training."— Presentation transcript:

1 2012 Annual Biosafety & Chemical Safety Training

2 Topics Emergency Response in Laboratories Chemical /General Lab Safety Biosafety and Bloodborne Pathogens Working with the IBC

3 Emergency Response in Laboratories

4 Lab Accidents Happen Chemical splash to eyes Chemical burns to skin Thermal burns Infectious exposures Needle punctures Lacerations Trip/slips/falls

5 Chemical Splash to Eyes at least 15 minutes Immediately flush with water for at least 15 minutes (remove contacts) Report to the Emergency Room or call 3333 Keep all eye wash stations free of obstructions

6 Chemical Exposure to Skin at least 15 minutes Immediately drench with water for at least 15 minutes Remove affected clothing Report to Emergency Room or call 3333 Always keep shower stations free of obstructions

7 General Chemical Spill Procedure S S top & evaluate size & severity of spill P P rotect any coworkers at risk I I solate the spill by closing doors L L aunch response action (call x4220) L L ocate MSDS Remember….. S. P. I. L. L S. P. I. L. L

8 General Chemical Spill Procedure CALL x4220 Large Spills (greater than 1 liter) Any spilled quantity of Highly Hazardous Materials Acutely Toxic Materials Any spill you cannot clean up yourself Report all incidents to: PI/Lab Supervisor AO/R&D; R&D Safety Officer Employee Health/ER if injured

9 Natural Gas Leak Emergencies If you smell or suspect it… 1.If minor, try to shut off valve(s). 2.If strong or cannot shut off valve(s), cease all operations immediately and DO NOT operate electrical devices (DO NOT unplug/ plug or turn on/ off anything) 3.Evacuate the area (DO NOT use elevators) 4.From a safe location, contact Police Service at (305)

10 Chemical /General Lab Safety

11 OSHAs Lab Standard Chemical Hygiene Plan: Provides policies and procedures for working safely in labs Outlines the proper use of engineering controls (e.g. fume hoods) and personal protective equipment Lab-specific SOPs to minimize chemical exposures The Chemical Hygiene Plan is in each laboratory The Miami VAHS Chemical Hygiene Officer is the R&D Safety Officer.

12 Material Safety Data Sheets Allow employees to better understand how to safely work with hazardous chemicals. Allow employees to better understand how to safely work with hazardous chemicals. Contains key information for a specific chemical, including health hazard information and control measures such as proper personal protective equipment, physical hazards and properties, handling and storage requirements, and emergency procedures. Contains key information for a specific chemical, including health hazard information and control measures such as proper personal protective equipment, physical hazards and properties, handling and storage requirements, and emergency procedures. All lab personnel should know how to read a MSDS and should review the MSDSs for chemicals used or stored in their work area. This will help prevent accidents and increase safety awareness. All lab personnel should know how to read a MSDS and should review the MSDSs for chemicals used or stored in their work area. This will help prevent accidents and increase safety awareness.

13 Material Safety Data Sheets Information includes: Chemical Name/ Synonym Manufacturer/Distributor Ingredients Chemical Characteristics Flammability/Reactivity Exposure Limit /Controls Health Hazard Info Signs & Symptoms of Exposure Disposal Guidelines

14 Material Safety Data Sheets Each lab must maintain its own chemical inventory, and MSDSs (paper). You can access electronically via manufacturers website or via

15 Chemical Inventory Per VAs regulation, must be updated twice a year. Health; Flammable; Reactive; Contact; Radioactive; None All acronyms must be defined

16 Basic Chemical Hazard Classes oxidizer corrosiv e flammabl e ga s poison

17 Oxidizer Agents that react with reducible material to initiate or promote combustion Must not be stored with flammable solvents or other organic chemicals Common oxidizers: - Hydrogen peroxide (concentrated) - Many nitrates and nitrites - Sulfuric, nitric, and chromic acids

18 Corrosives Acids & Bases Tissue damage React violently with water and each other Do not store acids & bases together Separate oxidizing acids (nitric, sulfuric) from organic acids (acetic acid, phenol)

19 Flammable Liquids Liquid with a Flashpoint < 100 o F Volatile, evaporate quickly, may lead to explosive concentration in air Always store large quantities in rated flammable storage cabinets Only keep small amounts out on benchtops and shelves Do not store in refrigerators or freezers not specifically designed for flammable chemical storage

20 Gas Flammable gas (CO, H 2 ) Flashpoint < 140 o F Ignites easily, burns rapidly Non-flammable gas (including compressed gas) (N 2, CO 2 ) Purified gas in pressurized tank Tank rupture, toxic atmosphere, oxygen displacement Store upright, secure with a double chain to prevent falling

21 Explosive/Shock Sensitive Thermodynamically unstable material Explosion caused by shock or chemical reaction Picric acid, 2,4-dinitro-phenol, organic azides Store minimum quantities; discard before expiration date

22 Toxics/Poisons/Carcinogens Harmful or fatal if inhaled, ingested, absorbed (Nicotine, Pesticides, Osmium Tetroxide) Carcinogen refers to any substance that is an agent directly involved in the promotion of cancer or in the increase of its propagation. (Dimethyl sulfate, Chloroform, Formaldehyde/ Formalin/Paraformaldehyde )

23 Chemical Exposure Pathways Inhalation – Chemical vapors or dust Ingestion – Hand hygiene is important! Injection – Needlesticks or broken glass Absorption – Direct skin contact due to not wearing gloves, or wearing inappropriate gloves Inhalation and absorption are most common

24 Chemical Fume Hoods Primary control for inhalation protection Must limit storage & clutter for proper functioning Keep sash at or below recommended level Perform all work 6 inside the hood Keep lab doors closed to ensure negative room pressure to the corridor and proper air flow into the hood.

25 Chemical Resistant Gloves Glove selection is important: – Variety of glove materials are available – No universal glove that is appropriate for all chemicals – All chemicals eventually penetrate all common glove materials – Gloves may appear to be intact even when they are not

26 Chemical Resistant Gloves Poor protection against many non- aqueous hazardous chemicals Many people have allergies to latex Latex is a sensitizer, meaning that repeated contact may cause you to develop an allergy Latex powder can trigger allergic reactions in your fellow employees Latex Gloves are strongly discouraged

27 Chemical Resistant Gloves Nitrile gloves most commonly used in research labs Pros: Low cost, good dexterity, resistance to wide range of chemicals (acids, bases, aliphatic solvents), resistant to abrasion and puncture Cons: Poor protection against acetone, acetonitrile, tetrahydrofuran, halogens, phenol (seek alternative gloves, such as butyl rubber)

28 ONE GLOVE REMINDER If transporting any hazardous materials outside the lab: HazMat = Glove Common Surfaces = No Glove Gloves should never touch door knobs, elevator buttons, etc.

29 Eye & Face Protection Types of protection: Safety Glasses Safety Goggles – better protection Face Shield – for splash hazards Prescription glasses do not qualify! Glass lenses, no side shields Contact Lenses are discouraged in labs Can react to chemical exposure and can interfere with eye washing

30 No sandals or open- toed shoes in the laboratory

31 No food or beverages in the lab! Imminent health hazard OSHA regulation COMMON SENSE! Throw food wrappers away before you enter the lab.

32 Chemical & Hazardous Material Storage Keep volumes to a minimum Avoid ordering more than a few weeks supply at a time Do not store chemicals on high shelves, under sinks, or anywhere else awkward to reach Use carts or bottle carriers to transport bottles between labs

33 Flammable & Combustible Chemicals Never order in bulk containers Prefer 1 gallon size limit. Labs with 10+ gallons need flammable storage cabinet

34 Segregate Incompatibles Separate Acids and Caustics Separate Organic and Mineral acids Separate Oxidizers from all Organics Keep flammables in rated cabinets and away from any ignition sources No combustibles (cardboard, paper) in or on your flammable storage cabinets

35 Peroxide-Forming Chemicals Repeated exposure to air causes peroxide buildup Unstable, potentially explosive VERY dangerous, very expensive ($5-10k) to dispose VERY dangerous, very expensive ($5-10k) to dispose Monitor expiration dates, order in small quantities ethyl ether picric acid 1,4-dioxane isopropyl ether

36 Biosafety and Bloodborne Pathogens

37 Biosafety in the Laboratory Primary goal of biosafety is to reduce or eliminate exposures to potentially hazardous biological agents through the use of containment. Biosafety guidelines are designed to protect both laboratory personnel and the community and environment outside of the laboratory.

38 Risk Groups Biological agents are categorized in Risk Groups (RG) based on their relative risk Four RGs - RG1 to RG4

39 Biosafety Level (BSL) Prescribe procedures and levels of containments for the particular agent used Four BSLs - BSL-1 to BSL-4 based on risk assessment

40 BSL-1 containment is typically required for lab work involving: Biological agents that meet the definition of RG1 (i.e. not associated with disease in healthy adult humans); Biological materials not suspected of containing RG2 or higher in a quantity or form that may cause human disease (e.g., many soils and nonprimate animal cells); Biological agents or materials not characterized by the supplier as RG2 or higher; Transgenic or wild-type lab animals (e.g., rodents) that can be contained and are: – Free of zoonotic disease – Not infected with, implanted with, or containing RG2 or higher agents or materials

41 BSL-2 containment is typically required for lab work involving: Biological agents/materials categorized as RG2 or may contain RG2 agents; Bloodborne pathogen materials (e.g., human blood, tissues or cells) Nonhuman primate tissues or cells; Lab animals (e.g., rodents) infected with, implanted with, or containing RG2 agents or materials (e.g., infected with a human pathogen or containing a xenotransplant of human cells) Tissues or cells potentially containing an RG2 agent

42 Three Components of Biosafety Laboratory practices and techniques – Primary goal is to minimize exposures to biological agents from inhalation, ingestion, or skin contact – Standard microbiological techniques applied Safety equipment – Primary barriers act as a first line of defense against exposures to hazardous agents (BSCs, PPE) Facility design – Secondary barriers protect laboratory workers inside the laboratory and the community outside of the laboratory and varies depending on the biosafety level (limiting access, autoclaves for decontamination, handwashing facilities)

43 Biosafety Level 1 Practices Access does not have to be restricted; however, doors cannot be propped open Handwashing after glove removal and prior to leaving laboratory Eating, drinking, smoking and applying cosmetics prohibited Mouth pipetting prohibited Sharps handled safely Work surfaces decontaminated Decontamination of biological waste

44 Biosafety Level 1: Safety Equipment Laboratory coats Gloves Eyewear protection for splashes and spills Closed-toe shoes Special containment equipment or facility design is not required, but may be used as determined by a risk assessment.

45 Biosafety Level 2 Practices (BSL-1 Practices Plus): Personnel have specific training in handling pathogenic agents Access to the laboratory is restricted when work is being conducted. Biohazard signs must be posted. Laboratory personnel must be provided with medical surveillance and offered appropriate immunization.

46 Biosafety Level 2: Safety Equipment Biological safety cabinet (BSC) needed when aerosols and splashes will be generated Centrifuge safety cups Face protection worn to minimize exposures to splashes and aerosols outside BSC Protective clothing worn only in laboratory Gloves worn when handling infectious materials or contaminated equipment Decontamination procedures

47 Biological Safety Cabinets Class II Type A2 is the most common type Optimal performance: n grills free of clutter n sash as low as possible n no open flame allowed inside BSC Not designed to work with volatile chemicals Annual certification is mandatory

48 Biological Safety Cabinets CleanDirty

49 UV lamps are not required or recommended in BSCs. If operated properly, BSCs do not need UV lights. If installed UV lamps must be: Cleaned weekly to remove dirt and dust (they block germicidal effectiveness of UV light) Checked periodically to ensure the appropriate intensity of UV light is being emitted Turned off when the room is occupied to protect eyes and skin from UV exposure **can burn the cornea and cause skin cancer Ultraviolet Lamps in BSCs

50 Ideal Vacuum Line Protection

51 Biohazardous Waste Containers

52 Sharps Precautions Never break, bend, resheath or reuse syringes or needles Always dispose of sharps in approved sharps containers

53 Bloodborne Pathogens (BBPs) OSHA Bloodborne Pathogens Standard (29 CFR ) Standard is designed to protect workers from health hazards associated with occupational exposures to blood and other potentially infectious material (OPIM) HIV, HBV, HCV, and SIV Biological agents that cause: n syphilis n malaria n viral hemorrhagic fever

54 Blood is a Potentially Infectious Material (PIM) OSHAs definition of blood includes: n Human and nonhuman primate blood n Human and nonhuman primate blood products n Human and nonhuman primate blood components such as plasma and platelets

55 Other Potentially Infectious Material (OPIM) Human body fluids including: n semen n vaginal secretions n breast milk n cerebrospinal, pleural, synovial, pericardial, peritoneal, and amniotic fluids n any body fluid visibly contaminated with blood Unfixed human and nonhuman primate tissues or organs Human and nonhuman primate cells or cell lines

56 Other Potentially Infectious Material Is Not: Tears Sweat Saliva Urine Unless visibly contaminated with blood

57 OSHA BBP Standard Written Exposure Control Plan Identification of all job titles with potential exposures to PIM Control of exposures to PIM Hepatitis B vaccination offered Communication of potential hazards to workers Records maintained

58 Required Labeling OSHA Standard requires that the biohazard symbol be affixed to any equipment that comes into contact with biohazard agents: n containers of regulated waste n Refrigerators/freezers n incubators n containers used to ship, store, or transport blood and OPIM

59 Bloodborne Pathogen Spills Alert people in immediate area of spill and post signs to keep people out of the area to prevent spread of the contamination. Wear the appropriate PPE to clean up spill: disposable gloves, lab coat, fluid resistant surgical mask, and safety glasses or goggles. Pick up sharps with tongs/forceps and discard into sharps container. Cover spill with wipes or paper towel. Spray spill with a freshly prepared 1:10 dilution of bleach and allow it to stand for at least 20 minutes. Wipe up the spill with paper towels, working from the edge toward the center. Wipe the area again with fresh paper towels soaked in disinfectant. Place towels in a double-lined biological waste box.

60 Disinfection and Decontamination Methods Alcohols (70 % isopropyl or ethyl alcohol solution) are effective for routine disinfection of surfaces Sodium hypochlorite (Bleach), 1:10 bleach dilution should be used for liquid biological waste decontamination Autoclaves

61 What If A BBP Exposure Occurs? Immediately treat the exposure site n Needlesticks and cuts should be washed with soap and water, apply direct pressure if needed n Splashes to the nose, mouth, or skin require flushing with water or saline for 3-5 minutes n Splashes to eyes: remove contact lens immediately if worn, go to eyewash station and immediately rinse eye(s) and inner surface(s) of eyelid(s) with water continuously for 15 minutes n Go to the Emergency Room n Notify your Supervisor n Notify the Research Safety Officer and Research Administration (Required by ORO)

62 Working with the IBC NIH requires all labs working with recombinant DNA to register with their local IBC Recombinant DNA (rDNA) – DNA constructed in vitro from separate DNA segments that can replicate and/or express a biologically active polynucleotide or polypeptide in vivo – Synthetic DNA that has the potential of generating a hazardous product in vivo UMs IBC policy requires that any recombinant work, exempt or non- exempt, be registered with the IBC. (

63 Working with the IBC When filling out the IBC forms make sure you pay special attention to: 1.What section of the NIH Guideline applies to you 2.Risk Groups and Biosafety Levels 3.Be as specific as possible in regards to your procedures, hosts, vectors, and genes 4.Always include the vector maps of your viral constructs Call:

64 Working with the IBC When completing IBC forms always include the vector maps Plasmid cloning vector in E. ColiLentiviral vectorAdeno-asssociated viral vector

65 Summary of NIH Guidelines Section III-C: Experiments that require IBC, and Institutional Review Board (IRB) approvals, and Recombinant DNA Advisory Committee (RAC) review before initiation. – Experiments involving the transfer of rDNA into human subjects. Section III-D: Experiments that require IBC approval before initiation. – Experiments involving the introduction of recombinant DNA into RG2, RG3 or RG4 agents for use in animal experiments. – Using adenovirus, adenovirus-luciferase or adeno-associated virus to transfect cells – Cloning a pathogen into a lower eukaryotic or prokarytoic cell – Insertion of KSHV (Kaposis sarcoma-associated herpesvirus) or RRV (Rhesus Rhadinovirus) genes into defective lentiviral vectors – Creation of transgenic animals or knockout animals that leave genetic material in the animal as part of the silencing of gene. (purchase or transfer of previously created transgenic rodents is exempt)

66 Summary of NIH Guidelines Section III-E: Experiments that require IBC notice simultaneously with initiation. – Experiments in which all components are derived from non-pathogenic prokaryotes or non-pathogenic lower eukaryotes – rDNA with less than 2/3 of a eukaryotic viral genome (and no helper virus) used exclusively in tissue culture (inserting DNA sequence that encode reporters that are measured (lacZ, luciferase, eGFP, etc)) Section III-F: Experiments that are exempt from NIH Guidelines. However, registration with the UM IBC is required. – rDNA molecules not in organisms or viruses – rDNA that cannot replicate or express in vivo – rDNA molecules that consist entirely of DNA segments from a single nonchromosomal or viral DNA source

67 More Questions... Reference materials (BMBL, NIH Guidelines) H&S Office x4220 Research Safety Officer x4436 Research Office x3179 Lab specific information should be obtained from lab supervisor or Principal Investigator

68 Things to Note No one should enter the lab areas without authorization. Need a VA Badge. Visitors must be accompanied by a VA employee at all times. Contractors must sign in with the Main Research Office prior to working. All unknown personnel entering a research area must be challenged – Example – lost patients, visitors without escorts – Can I help you? – What are you doing here? – Cant access this area without authorization All lab door should be closed and locked at all times. – Closed for negative pressure – Locked for security – theft is a big issue/concern Computer must be locked when you are not at them. Remove all copies from printers immediately regardless of whether you consider it to be sensitive information

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