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Biosafety in Research Laboratories

Definitions•Biohazard

•An agent of biological origin that has the capacity to produce negative effects on humans, plants or animals.

•Biosafety•The application of: lab practices and procedures, specific lab facilities and safety equipment to protect against exposure to potentially infectious material

What is Biohazardous Material?

BacteriaVirusesParasitesFungiHuman and Non-Human Primate MaterialRecombinant DNAAnimalsBiological Toxins

How Can You Be Exposed? Inhalation -80% of lab acquired infections (LAI)

Perform experiments in biological safety cabinets Use respiratory protection

Remaining 20% LAI are a combination of: Percutaneous

Avoid sharps when possible, use plastic pipettes and disposable glassware

Mucous Membrane (eyes, nose and mouth) Wash hands often, avoid touching face

Ingestion Wash hands often, No eating, drinking, smoking or applying

cosmetics

All exposures must be reported to the PI, Occ. Health, Biosafety

How Can You Protect Yourself?

Administrative Controls Use attenuated or non-virulent strains, require restraint

during animal injection, avoid sharps and glassware, require decontamination, Standard Operating Procedures, training

Engineering Controls Biosafety Cabinets, Sealed Rotors/Safety Cups, Pipette

Aids, Safety Enclosures for Cell SortersPersonal Protective Equipment

Lab Coat / gown, gloves, goggles, facial protection Close toed shoes

Guidelines and Regulations• Biosafety in Microbiological and

Biomedical Laboratories (CDC)• NIH Guidelines for Research

Involving Recombinant DNA Molecules (NIH)

• OSHA Bloodborne Pathogens Standard (OSHA)

• Committee on Microbiological Safety (COMS)

Why Biosafety Guidelines?

Protection• Self, Co-workers, Lab support personnel• Emergency Responders • Experiment and Materials • Environment

Compliance with NIH, CDC, OSHA, City of Boston, and Town of Belmont

Safety Equipment (Primary Barriers)

• Biosafety cabinets (BSCs)• Safety centrifuge cups and rotors• Pipetting Devices • Vacuum line HEPA Filters• Personal protective clothing

• Gloves (remember the one glove rule)

• Gowns/Lab coats (should remain inside the lab)

• Eye and face protection

• Close toed shoes

Biological Safety CabinetsMust be Certified Annually

• Air Systems Technologies (508) 427 5558

BSC Provide:• Product Protection• Personnel Protection• Environmental Protection

Use biosafety cabinets (class II) for work with infectious agents involving:

• Aerosols and splashes• Large volumes, High concentrations • BL2 materials

Biological Safety Cabinets Protection against particulates only No gas or vapor protection Air flow is easily disrupted

• Don’t cover the grill• Move arms slowly in and out of cabinet• Walk slowly

70% return 30% exhausted

Centrifuges

Safety Cups and RotorsLoad and unload safety cups inside the BSC

Vacuum Line HEPA Filter Protection

Placed in between the vacuum system and the aspiration flasks

Biosafety LevelsBSL-1: agents not known to cause disease (in

healthy adults)BSL-2: agents associated with diseaseBSL-3: indigenous/exotic agents

associated with human disease and with potential for aerosol

transmissionBSL-4: dangerous/exotic agents of life

threatening nature

Biosafety LevelsBSL-2S (with Stipulation)

• Harvard classification not recognized nationally

• Work can be done inside a BL2 structural facility while practicing certain BL3 practices and procedures

– disposable sleeve covers or gowns,.....

• Stipulated by the Institutional Biosafety Committee (COMS)

Biosafety Level 1Immunocompromised workers are at risk

Standard Operating Procedures Minimize splashes and aerosols PPE Decontaminate work surfaces daily Decontaminate wastes Maintain insect & rodent control program

Examples:• Adeno associated virus• Murine leukemia virus• Non pathogenic E. coli• Animal tissue and cells (other than NHP)

Biosafety Level 1 Laboratory Facilities (Secondary

Barriers)

Biosafety Level 2• Infectious dose will vary based on:

• Agent

• Exposure time

• Exposure route

• Immunization or antibiotic treatment may be available• Extreme precaution with contaminated needles or sharp instruments

• Examples:• Hepatitis B Virus

• Influenza

• Salmonella

Biosafety Level 2

Biosafety Level 2 Signs• Policies and

procedures for entry• Biohazard warning

signs Specific hazard training with

annual updates

Biosafety Level 2Laboratory Facilities (Secondary Barriers)

Hand washing sink located near laboratory exit

Autoclave available Eyewash/Shower station

available Biosafety Cabinets

Biosafety Level 2 Laboratory Facilities (Secondary Barriers)

Special Practices

Use leak-proof transport containers

Biological Spill Kits

• Location• Inside Tissue Culture Rooms

• Contents• Gloves, Goggles/shield, N95 Respirator, Lab coat/gown• Absorbent material• Bleach or suitable disinfectant• Tongs and dust pan

• Spill Procedures• Place absorbent material over spill• Pour disinfectant around outer edges of absorbent ending in the middle• Wait for inactivation of bio-hazardous material• Properly dispose of all materials in biohazard box

Biohazardous Waste Includes...

Blood and Blood ProductsCultures/Stocks of Infectious Agents and

Associated BiologicalsAnimal Carcasses and BeddingSharpsGlass / Volumetric PipettesPrimary and Established Human and Animal

Cell Lines

BL1 and BL2 Waste Disposal Procedures

Liquid: chemically treated prior to drain disposal 10% (total volume) sodium hypochlorite solution (Bleach; Hg

free) for 20-30 minutes

Solid: BL1 and BL2 non recombinant waste

• Dispose of directly into red plastic waste containers or cardboard boxes lined with red biohazard bags

Recombinant DNA• Any viable organism containing recombinant DNA must be rendered

noninfectious prior to disposal

• Steam sterilization or chemical inactivation

BL2S Waste Disposal

• Liquid• Inactivate all liquid waste with Vesphene then

autoclave

• Solid• Autoclave all solid waste

Avoid using Sharps and Glass if possible

Sharps Disposal

All sharps should be disposed of directly into an approved sharps container immediately after use

When the sharps container is 2/3 full: Place in biohazard waste container Arrange for pick up from Building Services (x2656)

COMS• Committee on Microbiological Safety

• The MGH Institutional Biosafety Committee (IBC)

• What research needs COMS approval Infectious microorganisms, in-vitro or in-vivo Recombinant DNA materials Human studies involving rDNA (Human Gene Transfer studies) Animal to human transplants (xenotransplantation) Primate tissues Dual use agents

All COMS approved research is subject to an annual biosafety inspection

The Forms: http://www.hms.harvard.edu/orsp/coms/

OSHA Bloodborne Pathogens Training

Bloodborne Pathogens

What are the three bloodborne

pathogens you need to be

concerned about and what is the

risk of transmission each?

BLOODBORNE PATHOGENS

HEPATITIS B (HBV) 30%

HEPATITIS C (HCV) 3 %

HIV 0.3%

Bloodborne Pathogens

Hepatitis B the most highly transmissible but greatly reduced by vaccine use!!

Hepatitis C No vaccine, but early identification and

treatment offers some hope

HIV Early treatment greatly reduces the risk

Exposure Prevention strategies

VaccinationWork Practices/Standard PrecautionsPPE/Engineering ControlsExposure responseEducation/polices

Hepatitis B vaccineAvailable from Occupational Health Service

(6-2217)Must be offered to all employees with

exposure riskA 3 shot series- must completeAntibody status checked after series

completedNo need for routine “boosters”

Work Practice ControlsStandard Precautions - Treat all blood and body fluids as infectious! - Gloves for contact with blood, body fluids, moist mucous membranes, non-intact skin, secretions and contaminated items - Other Personal Protective Equipment (PPE) as needed to suit the task you are performing - Remove PPE when task completed and moving to a clean area - Disinfect hands after contact with blood/body fluids/tissue, even if gloves are used - Disinfect equipment/surfaces per policy

PPE/Engineering Controls

Gowns, gloves, face shields, splashguards etc.

LabelingBiohazardous Waste DisposalSafety DevicesSharps Disposal

Sharps Containers

Post Exposure ResponseHow can you be exposed? -Percutaneous -Mucous membrane -Broken Skin

- Animal Bites Immediate Response -First Aid- wash areaReporting- Occ. Health (x2438) or call operator

(x2000), available 24/7-All animal bites must be reported to Occ. Health

Follow-up required

Policies and Procedures

Bloodborne Pathogens Exposure Control Plan- located in the Infection Control Manual

http://library.partners.org/MGH1/webserver/custom/trov

Departmental PoliciesAnnual Education