Biosafety Level 2 (BSL-2) Safety Guidelines

Biosafety Level 2 (BSL-2) Safety Guidelines

BSL-2 builds upon BSL-1. If you work in a lab that is designated a BSL-2, the microbes used pose moderate hazards to laboratory staff and the environment. The microbes are typically

indigenous and associated with diseases of varying severity. An example of a microbe that is typically worked with at a BSL-2 laboratory is Staphylococcus aureus.

In addition to BSL-1 considerations, BSL-2 laboratories have the following containment requirements:

Laboratory Practices

 Access to the laboratory is restricted when work is being conducted.

 Individuals must wash their hands: (a) after working with potentially hazardous

materials, recombinant materials and animals; (b) after removing gloves; and (c) before leaving the laboratory.

 Eating, drinking, smoking, handling contact lenses and applying cosmetics are not permitted in laboratory areas.

 Mouth pipetting is prohibited; mechanical pipetting devices must be used.

 Decontaminate all work surfaces after completion of work and after any spill or splash of potentially infectious or viable recombinant material with appropriate disinfectant.

o Suitable disinfectants include: 10% bleach solution, 70% ethanol solution or 2%

wescodyne solution.

 Decontaminate all cultures, stocks, and other potentially infectious or recombinant materials before disposal using an effective method.

Waste Disposal

 Liquid Waste – All liquid biohazard waste must be disinfected with 10% bleach solution, 70% ethanol solution or 2% wescodyne solution.

o Once the waste has been disinfected, flush it down the sink and allow the cold water to run for a period of time (at least 5 minutes).

 Solid Waste –All solid biohazard waste must be placed in a red or orange bag stored in an impervious container with a biohazard label on it; all solid biohazard waste must be autoclaved.

o Once the waste is autoclaved, mark the autoclaved bag with an “Autoclaved/

Decontaminated" sticker.

o Place the autoclaved bag into a black opaque bag and fully seal the bag.

o The autoclaved waste can be thrown into the regular trash.

 Sharps – All sharps must be placed in an approved sharps container.

o For uncontaminated or biologically contaminated sharps, contact Environmental Health & Safety (x6640) to arrange for a pick-up and delivery to Canton-Potsdam Hospital for disposal.

BLS-4 BSL-3 BSL-2 BSL-1

 For additional information on biohazard waste disposal, refer to Appendix I.

Safety Equipment

 Appropriate personal protective equipment (PPE) must be worn, including lab coats and gloves. Eye protection and face shields should also be worn, as needed.

 All procedures must be performed within a biological safety cabinet (BSC) .

 For additional information on safe use and maintenance of BSCs, refer to Appendix II.

Facility Construction

 The laboratory must have self-closing doors.

 A sink must be readily available for hand washing.

 An emergency eyewash station must be readily available for accidental exposure.

 A Clarkson University emergency contact sign must be posted on the outside of the door indicating that it is a BSL-2 designated lab by writing “2” in the Biohazard Level box.

Training

 Laboratory personnel must have documented specific training in the procedures

conducted in the laboratory regarding individual responsibilities, necessary precautions to prevent exposures and exposure evaluation procedures.

 Personnel must receive annual updates or additional training when procedural or policy changes occur.

Appendix I: Biohazardous Waste (BSL-1 and BSL-2) Disposal for Research & Teaching Laboratories

Examples of Materials Container Requirements Disposal Requirements Solid Waste All contaminated gloves,

paper towels, gauze, Kimwipes, absorbents, Petri dishes, culture vials, plastic vacutainers, lab equipment

 Must be leak– and puncture proof

 Line with a red or orange biohazard bag

 Lid must be in place when waste is not being added to container

 Label with

“BIOHAZARD” sticker

 Keep container and lid clean at all times.

 Maintain access to the container - do not store items on lid

 Remove all liquid from solid materials - liquids must not be placed in the bags as they can easily leak.

 Place all solid biohazard waste in a red or orange bag.

− Red and orange bags must be stored in impervious containers to prevent leaking or contamination.

− It is a recommended practice to double bag the hazardous material, but is not required.

 All solid biohazardous waste must be autoclaved.

 Prior to autoclaving, crisscross the bag's biohazard symbol and/or markings with heat sensitive autoclave tape.

 Ensure the autoclave is set for the appropriate time - setting the autoclave for too short a period may not properly disinfect the material.

 Once the waste is autoclaved, mark the autoclaved bag with an

“Autoclaved/Decontaminated" sticker.

 Place the autoclaved bag into a black opaque bag and fully seal the bag.

 The autoclaved waste can be thrown into the regular trash.

Liquid Waste Any liquid from Petri dishes, culture vials, vacutainers, lab equipment, etc.

N/A  Liquid waste must be separated from solid waste

 Liquid waste must be decontaminated with an appropriate disinfectant

− 10% bleach solution

− 70% ethanol solution

− 2% westcodyne solution

 Flush the disinfected material down the sink, allowing the cold water to run for a period of time (at least 5 minutes).

 Do not flush non-aqueous solutions, such as agarose gels or broths, down the drain as they will clog the pipes.

Examples of Materials Container Requirements Disposal Requirements Sharps All needles, syringes,

scalpels, razor blades, pipette tips, Pasteur pipettes, glass vacutainers, capillary tubes, slides and cover slips, contaminated broken glassware

 Must be rigid, leak- proof, and puncture- proof

 Single use only

 Keep baffle in place

 Label with “BIOHAZARD SHARPS” sticker

 All sharps must be placed in appropriate sharps container.

 Once the container is ¾ full, close the top of the container.

 If the sharps are chemically contaminated, place a yellow

“Hazardous Waste” label on the container and complete the label. Cross out the biohazard symbol on the container. Contact the Chemical Stockroom for a hazardous waste pickup at x2338.

 If the sharps are radioactive, cross out the biohazard symbol and place a radioactive material sticker on the container. Contact the Radiation Safety Officer to request a waste pickup at x6640.

 If the sharps are biohazardous or not contaminated,

arrangements should be made with the Environmental Health &

Safety Manager (x6640) to have the waste transported to the Canton-Potsdam Hospital.

Pathological Animal carcasses suspected to be or potentially infected;

Recognizable human tissue, organs and body parts

 Must be leak-proof and puncture-proof

 Lid must be in place when waste is not being added to container

 Label with

“BIOHAZARD” and

“PATHOLOGICAL WASTE” stickers

 Waste must be placed in red or orange biohazard waste bags.

 If waste is chemically contaminated (ex. preserved in

formaldehyde or other similar substance), place a yellow hazard waste sticker on the bag. Contact the Chemical Stockroom for a waste pick-up at x2338.

 If waste is not chemically contaminated, arrangements should be made with the Environmental Health & Safety Manager (x6640) to have the waste transported to the Canton-Potsdam Hospital.

Appendix II: Safe Use & Maintenance of Biological Safety Cabinets (BSCs)

 A biological safety cabinet is specially designed and constructed to offer protection to both the worker and the biological materials. It’s used as the primary barrier against exposure to biohazardous or infectious agents, as it surrounds the immediate workspace.

 A conventional fume hood should not be used for work with viable biological agents.

 A biological safety cabinet should generally not be used for work with hazardous chemicals.

 Most biological safety cabinets exhaust the contaminated air through high efficiency particulate air (HEPA) filters back into the laboratory. This type of filter will not contain most hazardous materials, particularly gases, fumes or vapors.

 Even when connected to the building exhaust system, a ducted biological safety cabinet may not achieve a face velocity of 95 - 125 feet per minute, making it inappropriate for use with hazardous chemicals.

 It is the department’s responsibility to hire an outside contractor to inspect their biological safety cabinets on an annual basis.

Working in a Biological Safety Cabinet 1. Plan

 Thoroughly understand procedures and equipment required before beginning work.

 Arrange for minimal disruptions.

2. Turn on Cabinet

 Allow the cabinet to operate unobstructed for at least fifteen (15) minutes.

3. Disinfect

 Wipe down the interior surfaces of the cabinet with 10% bleach followed by 70% ethanol and allow it to dry.

4. Prepare Work

 Place your supplies into the cabinet as far away from the sash as possible.

 All operations should be performed on the work surface at least four (4) inches from the inside edge of the front grill.

 Do not use open flames in the cabinet.

5. Clean Up

 Wipe down all materials with disinfectant before removing from the BSC.

 After the cabinet is emptied, wipe down the interior cabinet surfaces with disinfectant.

 Allow the BSC to run for fifteen (15) minutes before turning it off.

 Remove gloves and lab coat carefully and wash hands.