2012 Annual Biosafety & Chemical Safety Training

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

Emergency Response in Laboratories Regardless of the workplace, emergencies will occur. Laboratories, even well controlled, are not immune to accidents and emergencies.

Lab Accidents Happen Chemical splash to eyes Chemical burns to skin Thermal burns Infectious exposures Needle punctures Lacerations Trip/slips/falls Whether by carelessness or naivety, or by way of “unforeseen circumstances” such as a loss of electrical power, it is a good bet that every researcher will eventually experience an emergency sometime during his or her career. Whatever the reason is, we have to understand that working in a laboratory comes with some inherent risks and that safety to personnel should be our number one concern. The severity of the outcome depends on what’s involved and how effective we are at responding to it.

Chemical Splash to Eyes 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 Depending on the chemical and degree of exposure, the potential for injury due to chemical splash to the eyes ranges from temporary redness and irritation to blindness. So what should you do if there is a chemical splash to the eyes?

Chemical Exposure to Skin 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 Skin exposure to chemicals in the workplace is a significant problem in the US.  Most chemicals are readily absorbed through the skin and can cause mild to serious health effects. Absorption of chemicals through the skin can occur without being noticed by the worker. So what should if there is a chemical exposure to the skin?

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

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

Natural Gas Leak Emergencies If you smell or suspect it… If minor, try to shut off valve(s). 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) Evacuate the area (DO NOT use elevators) From a safe location, contact Police Service at (305) 575-3333 Natural gas in the laboratory is used mainly with Bunsen burner to produce a single open gas flame which is used often for heating and sterilization. A gas leak refers to the seeping of natural gas, frequently through a pipe, which can endanger the lives of those in the area. While the fumes may cause health problems, the larger problem is the potential for an explosion because natural gas is highly flammable. 

Chemical /General Lab Safety By the very nature of laboratory work , it can be a place of danger if proper common-sense precautions aren’t taken. Knowing the types of chemicals that you will be working with, what general safety guidelines to follow and what safety precautions to take will ensure a safe laboratory environment for both yourself and the people you may be working near.

OSHA’s 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.

Material Safety Data Sheets 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. All lab personnel should know how to read a MSDS and should review the MSDS’s for chemicals used or stored in their work area. This will help prevent accidents and increase safety awareness. 1st: Material Safety Data Sheet or MSDS. 2nd: MSDS are intended to give end users a general understanding on how to safely use the material. 3rd:Each MSDS contains health hazards, physical properties, fire fighting, and PPE recommendations. 4th: All lab personnel are required to have a general understanding of what recommendations are included in the MSDS for each hazardous material he/she uses in the lab. Doing so will help prevent accidents and increase safety awareness.

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 1st: MSDSs include: Chemical name, manufacturer, ingredients, chemical characteristics, flammability/reactivity, exposure limits/controls, health hazard info, signs & symptoms of exposure, disposal guidelines.

Material Safety Data Sheets Each lab must maintain its own chemical inventory, and MSDSs (paper). You can access electronically via manufacturer’s website or via http://vaww.ceosh.med.va.gov/ceosh/MSDS.shtml 1st: Each lab must maintain hard copies of each MSDS. MSDS is obtain at time of delivery or you can access the manufacturer’s website or via the following site: http://vaww.ceosh.med.va.gov/ceosh/MSDS.shtml

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

Basic Chemical Hazard Classes flammable oxidizer gas corrosive poison There are many types of chemical hazards. In this training, we will cover five basic chemical hazard classes, which are Flammable, Oxidizer, gas, corrosive, and poison (toxic).

Oxidizer - Hydrogen peroxide (concentrated) 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 1st: The first class we are to review is Oxidizers 2nd: Oxidizers are materials that Supply oxygen to fuel a fire 3rd: The primary concern is that oxidizers must not be stored with flammable solvents or other organic chemicals.

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) 1st: A corrosive substance is one that will destroy or damage another substance with which it comes in contact. The main hazards to people include damage to eyes, skin and tissue under the skin, but inhalation or ingestion of a corrosive substance can damage the respiratory tracts. Exposure results in chemical burn.

Flammable Liquids Liquid with a Flashpoint < 100oF 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 1st: Generally, a flammable liquid means a liquid which may easily catch fire (flash point of less than 100 degrees Fahrenheit). The flash point of a liquid is the lowest temperature at where the liquid will ignite.

Gas Flammable gas (CO, H2) Flashpoint < 140oF Ignites easily, burns rapidly Non-flammable gas (including compressed gas) (N2, CO2) Purified gas in pressurized tank Tank rupture, toxic atmosphere, oxygen displacement Store upright, secure with a double chain to prevent falling

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

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) 1st: In the context of biology, toxic/poison substances are substances that can cause disturbances to organism and can be harmful or fatal if inhaled, ingested or absorbed.

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 1st: Chemical exposure pathways include inhalation, ingestion, injection, and absorption. 2nd: The most common routes of exposures are inhalation and absorption, which is why proper skin protection by using appropriate gloves and using toxic chemicals inside of your chemical fume hood. Inhalation and absorption are most common

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. 1st: Chemical fume hoods are the primary control for inhalation protection and designated area for chemical use.

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 Since you may have potential direct skin contact with several hazardous materials, wearing skin protection is mandatory when using any hazardous material. Remember that: 1- no one glove protects against all chemicals. 2- chemicals will eventually penetrate all common gloves

Chemical Resistant Gloves Latex Gloves are strongly discouraged 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 Later gloves provide poor to no protection against many non-aqueous hazardous chemicals, the powder in latex gloves can cause allergic reaction. Therefore, latex gloves are strongly discouraged. It is recommended that lab personnel should use nitrile rubber gloves.

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)

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. One Glove Reminder: If you’re transporting any hazardous material outside of your lab, you should never touch door knobs, elevators buttons, or other common surface with your gloved hand.

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 1st: Due to the potential splash hazard to your eyes and face, eye and face protection is required when working with any hazardous material where the process puts you or someone else at risk of a splash. 2nd: Some common types of eye and face protection are: 1-Safety glasses- which are normally used for impact hazards, such as flying objects. 2- Safety or chemical goggles- these provide better protection for chemical splash hazards to your eye, not your face! 3- Faceshields- provides protection for splash hazards to face. 3rd: Remember that prescription glasses do not qualify as safety glasses! 4th: Also, the use of contact lenses in laboratories is discourage!

No sandals or open-toed shoes in the laboratory

No food or beverages in the lab! Imminent health hazard OSHA regulation COMMON SENSE! 1st: Although you may see it as a common sense, ingestion incidents from personnel eating inside of laboratory is still an issue. 2nd: OSHA’s lab standard prohibits it. 3rd: Remember to throw food and their wrappers before you enter a lab. Throw food wrappers away before you enter the lab.

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

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

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 To prevent chemical hazard incidents, lab personnel should be vigilante on how and where the chemicals are stored. Chemicals should be stored according to their compatibility. Segregating incompatible chemicals will go a long way in providing lab personnel a safe environment to work in.

Peroxide-Forming Chemicals Repeated exposure to air causes peroxide buildup Unstable, potentially explosive VERY dangerous, very expensive ($5-10k) to dispose Monitor expiration dates, order in small quantities ethyl ether picric acid 1,4-dioxane isopropyl ether Certain chemicals can form peroxide either upon aging, or upon contact with air or other substances. Some of the peroxides are shock sensitive and can explode if handled less than gingerly, or upon heating. 

Biosafety and Bloodborne Pathogens Biohazard Symbol: Developed as “universal symbol” to warn of dangers from infectious or potentially infectious agents Unique and unambiguous Quickly recognized Easily recalled

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.

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

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

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 The Biosafety level will be determined as part of the risk assessment.

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 For BSL2+: Adenoviral vectors, Adeno-Associated Virus (AAV), Lentiviral vectors, and Retroviral vectors

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)

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

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. 

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.

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

Biological Safety Cabinets Class II Type A2 is the most common type Optimal performance: grills free of clutter sash as low as possible no open flame allowed inside BSC Not designed to work with volatile chemicals Annual certification is mandatory The primary purpose of a BSC is to serve as the primary means to protect the laboratory worker and the surrounding environment from pathogens. There are several classes of BSC, but the most common type is Class II Type A2. Class II cabinets are ventilated cabinets that use HEPA filtration, laminar air flow, and containment to protect the operator, the product, and the environment from aerosols generated from microbiological or pharmaceutical procedures performed within the lab.

Biological Safety Cabinets Clean Dirty Equipment is laid out to not restrict airflow in the cabinet A typical layout for working “clean to dirty” within a Class II BSC. Clean cultures (left) can be inoculated (center); contaminated pipettes can be discarded in the shallow pan and other contaminated materials can be placed in the biohazard bag (right).  48

Ultraviolet Lamps in BSCs 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

Ideal Vacuum Line Protection

Biohazardous Waste Containers In the research lab or field environment, biohazard non-sharp waste includes any non-sharp item that is contaminated with human or animal diagnostic specimen material (i.e., body fluids, tissue debris), any microbiological culture material (including recombinant DNA). This type of waste must be collected for final treatment and disposal in a leak-proof container lined with a bag of moderate thickness to prevent punctures. The collection container must have a lid or other means of closure and the container must be labeled with the biohazard symbol regardless of the lab’s operating biosafety level.

Sharps Precautions Never break, bend, resheath or reuse syringes or needles Always dispose of sharps in approved sharps containers Sharps are objects that can cause skin punctures or cuts (percutaneous exposures) Examples include : hypodermic needles and syringes Pasteur pipettes (made of glass) scalpel blades and razor blades glass microscope slides microtome knives suture needles broken glass

Bloodborne Pathogens (BBPs) OSHA Bloodborne Pathogens Standard (29 CFR 1910.1030) 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: syphilis malaria viral hemorrhagic fever Bloodborne pathogens require implementation of biosafety level 2 (BL2) practices and containment

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

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

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

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

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

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.

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

What If A BBP Exposure Occurs? Immediately treat the exposure site Needlesticks and cuts should be washed with soap and water, apply direct pressure if needed Splashes to the nose, mouth, or skin require flushing with water or saline for 3-5 minutes 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 Go to the Emergency Room Notify your Supervisor Notify the Research Safety Officer and Research Administration (Required by ORO)

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 UM’s IBC policy requires that any recombinant work, exempt or non-exempt, be registered with the IBC. (http://uresearch.miami.edu/default.asp?p=177) Due to ORO site visit back in January of this year, some protocols were cited as not having gone through the proper IBC approval process. Since then, we have developed a memorandum of understanding with the University of Miami to utilize their IBC. Some of you have already been contacted by Katrina or Dr. Kapsalis regarding this. All forms required for IBC review are posted on the UM website: (http://uresearch.miami.edu/default.asp?p=177).

Working with the IBC When filling out the IBC forms make sure you pay special attention to: What section of the NIH Guideline applies to you Risk Groups and Biosafety Levels Be as specific as possible in regards to your procedures, hosts, vectors, and genes Always include the vector maps of your viral constructs What to include in these forms is the most common question. Since the reviewer will not have your entire protocol packet, it is important to include as much information as possible so that the reviewer knows what you proposed to do. As always, it is important to include maps of any vectors that you will be using. Call: 305-243-2311

Working with the IBC When completing IBC forms always include the vector maps A vector, or a specific plasmid that is used for generating recombinant DNA. IBC requires the vector maps to check to see if you are using the correct ones. Vectors come in a variety of forms, but they must have three sequences. 1.They need an origin of replication (oriR) site, which is a sequence that signals bacteria to replicate this plasmid whenever the bacteria replicate.   2.There must be an antibiotic resistance gene, which is important for selecting bacteria that have therecombinant plasmid. When a scientist grows the bacteria, he will add an antibiotic to kill all bacteria that do not have the plasmid. The antibiotic resistance gene will allow the bacteria that have the plasmid to stay alive. Score.   3.A multiple cloning site (MCS) is important for facilitating insertion of the target gene into the vector. Putting the MCS into a lacZ or ccdB gene allows selection for bacteria that have the target gene inserted into the vector. This is because the target gene will disrupt the lacZ or ccdB gene, and one can easily test the activity of either of these genes by looking for colonies on a plate that lack the relevant gene activity. Plasmid cloning vector in E. Coli Lentiviral vector Adeno-asssociated viral vector

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 (Kaposi’s 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) The NIH Guidelines has six sections (Sections IIIA-F) that covers a myriad of rDNA experiments. Most labs at the VA will be dealing with Sections IIIC-F.

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 A helper virus is a virus used when producing copies of a helper dependent viral vector which does not have the ability to replicate on its own. The helper virus is used to coinfect cells alongside the viral vector and provides the necessary enzymes for replication of the genome of the viral vector.

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

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? Can’t 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 Heads up – unannounced fire and safety drills will be conducted soon.