Basic Biosafety Principles EH&S Academy Brenda J. Wong, UCSD Biosafety Officer October 2009.

Basic Biosafety Principles

EH&S Academy

Brenda J. Wong, UCSD Biosafety Officer

October 2009

What is Biosafety?

Safety from exposure to Infectious Agents

Smallpox

What is Biosafety?

Sunday, Sep. 20, 2009

Did the Plague Kill Illinois Scientist?By AP

(AP / CHICAGO) — The University of Chicago Medical Center says the infection that killed a scientist may be connected to bacteria he researched that causes the plague.

The university said Saturday that its researcher studied the genetics of harmful bacteria including Yersinia pestis, which causes the illness. He died Sept. 13. His name and age haven't been

released

The medical center says the bacteria he worked with was a weakened strain that isn't known to cause illness in healthy adults. The strain was approved by the Centers for Disease Control and

Prevention for laboratory studies.

An autopsy found no obvious cause of death but did find the presence of the bacteria. More tests are planned. No other illnesses have been reported.

Biosafety in Various DisciplinesBiosafety is related to several fields

ECOLOGY: referring to imported life forms not indigenous to the region (Reggie the alligator)

AGRICULTURE: reducing the risk of alien viral or transgenic genes, or prions such as BSE/"MadCow“; reducing the risk of food bacterial contamination

MEDICINE: referring to organs or tissues from biological origin, or genetic therapy products, virus; levels of lab containment protocols BSL-1, 2, 3, 4 in rising order of danger

CHEMISTRY: i.e., nitrates in water, PCB levels affecting fertility

EXOBIOLOGY: i.e., NASA's policy for containing alien microbes that may exist on space samples - sometimes called "biosafety level 5"

Biosafety in Academic Research Research Universities: Promoting safe laboratory

practices, and procedures; proper use of containment equipment and facilities; provides advice on laboratory design and risk assesment of experiments involving infectious agents, rDNA in-vitro and in-vivo.

Bottom Line: Risk & Containment

Biohazard Symbol Charles Baldwin at

National Cancer Institute at NIH.

Symbol to be “memorable but meaningless” so it could be learned.

Blaze orange – most visible under harsh conditions

Biosafety Issues Laboratory Safety Bloodborne pathogens (BBP) Recombinant DNA (rDNA) Biological waste disposal Infectious substance and diagnostic specimen shipping

Biosafety Issues (con’t.)

Respiratory Protection Bioterrorism and Select agents Mold and indoor air quality Occupational safety and health in the use

of research animals Biohazards used in animal models

Biohazardous Materials Viruses Bacteria Fungi Chlamydiae/Rickettsiae Prions Recombinant DNA

Biohazardous materials Transgenic Plants, Animals and Insects

Transgenic Insects

Biohazardous Materials

Human and Primate Cells, Tissues, and Body Fluids

Brain Tissue from Demented Patients Viral Vectors

Replication deficient viruses

Biosafety In Microbiologicaland Biomedical Laboratories“BMBL” (acronym)

CDC/NIH PublicationSafety “Guidelines”Regulations of Institution receives NIH funding

Code of Practice and “Gold” Standard in Industry anlGold Standard

Clinical & Research Lab.Lab. Animal Facilities

Biosafety Concepts

HHS Publication No. (CDC) 93-8395

The New BMBL

Early print edition….

Emphasis on “Risk

& Containment”

The BMBL continues to be published by the CDC and the NIH 5th edition is now at the printers

http://www.cdc.gov/od/ohs/biosfty/bmbl5/bmbl5toc.htm

Biosafety ConceptsThe BMBL

Are the NIH Guidelines Optional?

“Guidelines” does not mean “optional”

They are a term and condition of NIH funding for recombinant DNA research.

From Kathryn Harris, NIH, OBA

Biosafety Concepts from the BMBL

Principles of Biosafety Practice and Procedures

Standard Practices Special Practices & Considerations

Safety Equipment Facility Design and Construction Increasing levels of protection

Principles of Biosafety

Biosafety Levels 1-4 (BSL) Increasing levels of employee and environmental protection Guidelines for working safely in research & medical

laboratory facilities

Animal Biosafety Levels 1- 4 (ABSL)

Laboratory animal facilities Animal models that support research Guidelines for working safely in animal research facilities

Biosafety ConceptsThe BMBL

(1) Standard Microbiological Practices Most important concept / Strict adherence Aware of potential hazard Trained & proficient in techniques Supervisors responsible for:

Appropriate Laboratory facilities Personnel & Training

Special practices & precautions Occupational Health Programs

Biosafety IssuesThe BMBL

(2) Safety Equipment Primary Containment Barrier Minimize exposure to hazard

Prevent contact / Contain aerosols Engineering controls/ equipment Personal Protective Equipment (PPE)

Gloves, gowns, Respirator, Face shield, Booties Biological Safety Cabinets Covered or ventilated animal cage systems

Biosafety ConceptsThe BMBL

(3) Facility Design and Construction

Secondary Barrier/ Engineering controls

Contributes to worker protection Protects outside the laboratory

Environment & Neighborhood Ex. Building & Lab design,

Ventilation, Autoclaves, Cage wash facilities, etc.

Laboratory Design

“Warehouse Type Lab”

Discussion

What are some of the negatives and positives of this open lab concept?

Biosafety Level-1 Concepts of Biosafety

Biosafety Level-1 (BSL-1 or ABSL-1) Well characterized agents Agents not known to cause disease (in healthy human

adults; now healthy immunocompetent adults)

Prophylactic treatment available Open bench procedures Animals in open cage system or open environment

(outdoors) Good laboratory practices

Risk Group 1 Agents E.coli K-12 Transgenic Plants Plasmids Fungi Mold Yeast

BSL-1 Practices

Bench-top work allowed Daily Decontamination Manual pipetting Required Handwashing Red bag waste Bio cabinet not required

(unless creating aerosols) 2˚ containment

Risk Group 2 Agents Human or Primate

Cells Herpes Simplex Virus Replication

Incompetent Attenuated Human Immunodeficiency Virus

Patient specimens

BSL-2 Practices Concepts of Biosafety

Practices & Procedures Agents associated w/ human disease Treatment for disease available Agent poses moderate hazard to personnel and

environment Direct contact or exposure Percutaneous exposure

Scratch, Puncture, Needle stick Mucus membrane exposure

Eyes, Mouth, open cut

BSL-2 Practices

Limited access to lab when work in progress

Daily decontamination Mechanical pipetting Labcoat, safety glasses

and gloves required Red bag & sharps

containers required

BSL-2 Practices (con’t)

Biohaz. Sign posted at entrance to lab

Label all equipment (incubators, freezers, etc.)

TC room – negative air flow Documented training Baseline serology or pre-

vaccination may be required

Risk Group 3 Agents Human

Immunodeficiency Virus

Mycobacterium tuberculosis

Coxiella burnetii

Biosafety Level 3Working in High Containment

Biosafety Level-3 (BSL-3 or ABSL-3)

Indigenous or exotic agents Aerosol transmission Serious health effects Treatment may or may not exist

BSL-3 Practices

Public access NOT permitted Daily decontamination after spill and

upon completion of experiment Autoclave required and waste is

disposed at the end of day Required foot activated handwashing

sink and controls No sharps unless absolutely necessary

BSL-3 Practices (con’t)

Aerosol minimization procedures required Wrap around disposable clothing is

required. Specialized equipment may be required depending upon procedures

Biohaz. Signs and labels posted Air flow from low hazard to high hazard

“Pressure Mapping”

BSL-3 Practices (con’t)

Bench top work not permitted Documented training and personnel competency

certification (for BSL-3 procedures) Baseline serology Spills – report immediately and treat accordingly Vaccinations/post exposure protocols and SOP’s,

Biosafety Manual, Biosafety Officer

UCSD’s BSL-3

Biosafety Level-4Working in High Containment

Biosafety Level-4 Builds on BSL-3/ ABSL-3 practices Maximum containment facilities Pressurized Containment Suite

BSL-3 + Class III Biosafety Cabinet Chemical decontamination showers Liquid effluent collection / decontamination No BSL-4 labs exist at UCSD

Biosafety Level 4 Lassa Fever Virus Ebola Hemmorrhagic

Fever Virus Marburg Virus Herpes B Virus

Biosafety ConceptsWorking in High Containment

Biosafety Level-4 (BSL-4 or ABSL-4) Dangerous/exotic agents Life threatening disease Aerosol transmission Agents of unknown risk of transmission or health affects No known treatment

Animal Biosafety Level-4Working in High Containment

General Good Lab Technique

Hygienic Practices No Smoking, Eating, Applying cosmetics, lip

balm, contacts Wash hands after procedures Decontaminate lab bench before and after

work

General Operational Practices Proper attire

Minimum – lab coat, safety glasses, gloves Plan your work

Know in advance what you are working with Read available resources (MSDS)

http://www.hc-sc.gc.ca/pphb-dgspsp/msds-ftss/index.html

Animal Containment Points

CDC - 1957CDC & UCSD - 2005

Courtesy of Paul Vinson, CDC

Discussion # 2 Based on what you know about Biosafety

Levels, Practices and Operational Controls, what are some discussion issues for conducting Biohazard risk assessments?

How do you approach risks when addressing a particular organism?

Risk Assessment

In-Vitro In-Vivo Human ClinicalTrial

Addressing Risk Assessments What is the organism? Is it Wild-type, attenuated, irradiated, or

chemically treated? Look at kill data or kill curves.

What is the max. concentration, volume, infectious dose?

What is the work space like? Aerosolizing procedures? How do they contain

their aerosols?

Risk Assessment, con’t Are personnel trained? Do

personnel understand the organism, infectious dose and symptoms?

What are their experimental procedures?

Will they be transporting the material? Shipping intra, inter-state or international?

Are they doing tissue culture? Do they have adequate

containment equipment?

Tom Pugh

Risk Assessment, Con’t Are they doing this

work in-vivo? Have you consulted and discussed this with the Vets and IACUC to determine special needs and housing?

Waste issues addressed?

Pregnancy issues with the organisms?

Risk Assessment, con’t Do they share their

Tissue Culture room? Do they have more than

1 Biosafety Cabinet? Occupational Health

informed and set up to receive patient or offer counseling?

Accidental Spills

Evacuate area, alert personnel and cordon off so that aerosols may settle Don PPE; Cover with paper towels and apply bleach (1 part bleach : 9 parts water Allow 15 – 20 min contact time Wipe up working towards center Use tongs if broken glass is involvedIs Recombinant DNA involved?

First Aid Measures

Splash to Eye or Needlestick Injury Rinse thoroughly for 15 minutes at the eyewash or

sink Call Occupational Medicine 619 471-9210 Call EH&S to report exposure – 858 534-5366

What Helps?

Resources UCSD Biosafety: http://blink.ucsd.edu/safety/research-

lab/biosafety/

Biological Safety MSDS: http://www.phac-aspc.gc.ca/msds-ftss/index-eng.php

NIH BMBL:

http://www.cdc.gov/od/ohs/biosfty/bmbl5/bmbl5toc.htm See Handout for additional resources..

Questions?

Brenda Wong

[email protected]

858 534-6059

Cell: 858 583-3277

Additional Resources:

Biosafety Resources.doc