Northern Michigan UniversityBiology Department

Biology Laboratory Safety

Biohazard Safety Module

This presentation is designed as a training course for those students who will be intentionally working with Biohazardous materials

Biohazards are defined as those materials or organisms that carry the possibility of causing disease

Recombinant DNA

Artificial Gene Transfer

Infectious Agents (includes attenuated lab & vaccine strains)

Biologically Derived Toxins

The NMU Institutional Biological Safety Committee oversees research and teaching that deals with…

Biohazardous Agents by Risk Groups (RG)

Agents: any infectious entity or any molecule produced by them capable of producing disease in humans

At NMU, we are approved to work with RG1 and RG2 agents for research purposes. However, students could be exposed to RG3 agents from the environment.

RG1 Agents that are not associated with disease in healthy adult humans

RG2 Agents associated with human disease that is rarely serious or forwhich preventive or therapeutic interventions are often available

RG3 Agents that are associated with serious or lethal human disease for which preventive or therapeutic interventions may be available. The agent is of low risk for the community

RG4 Agents that are likely to cause serious or lethal human disease for which preventive or therapeutic interventions are not usually available. The agent is of low risk for the community.

Selected agents

Examples of pathogens and toxins

E. coli nonpathogenic laboratory strainsSaccharomyces cereviseaeBacteriophages

RG1 group

RG2 group

E. coli, pathogenic strainsInfluenza virusPseudomonas aeruginosa

Examples of pathogens and toxins

Human immunodeficiency virusHistoplasma capsulatumMycobacterium tuberculosis

RG3 group

RG4 group

Ebola virusMarburg virus

Routes of transmission of infectious agents in the lab setting:

Oral Ingestion Eating, drinking, smoking Broken skin Needle-stick Cuts Scratches Bites

Mucosa Splash of liquids Handling eye contact lenses Applying make-up

Upper respiratory tract Inhalation of aerosols

Oral Ingestion

It can take fewer than 10 E. coli O157 H7 to infect a human!

NO drinking or eating in the lab!

International Biohazard Symbol

Universal Symbol

Communicates potential exposure

Red or orange

Any container with biohazard potential must be labeled with this symbol

Needles and syringes, scalpels, blades, broken glass, etc.

Glassware exposed to an infectious agent such as Pasteur pipettes, glass containers, etc.

Broken skin

SHARPS

Take care when dealing with…

Any potentially contaminated sharp should be disposed of in a Biohazard Sharps container

Sharp Containers for Biohazards

Sharp containers are red, display the International Biohazard Symbol are labeled Medical Waste or Infectious Waste are puncture proof

Mucosa Exposure Routes

Good safety practice!!

No!

Make-up

Contact lenses

Opening tubes CentrifugationFlaming to sterilize toolsVortexingPipettingElectroporationSonicationFlow cytometry

REMEMBER: An agent that is not normally airborne transmitted can be transmitted in the lab by aerosols

Upper respiratory tract exposure methods

Some activities that generate aerosols – take care!

Blood and Other Potentially Infectious Material (OPIM)

Follow "Universal Precautions" all human blood and certain human body fluids are

treated as if known to be infectious

Other Potential Sources of Infection

Environmental Samples: soil, plants, water samples, animals, etc.

Follow “NMU Precautions“all environmental samples are treated as potential

health hazards

Primary containment: protection of people & immediate laboratory environment

Containment of Biohazard Agents

The purpose of containment is to reduce or eliminate exposure of laboratory workers & the outside environment to hazardous agents

Secondary containment : protection of external environment

It is achieved by the use of Good Microbiological Techniques Safety Equipment

It is achieved by the use of Facility Design

Operational Practices

Primary containment

Protection of people and the immediate laboratory environment

Good Microbiological Techniques

Use proper Safety Equipment (Primary Barriers):

Engineering controls designed to remove or minimize exposures to hazardous biological materials: safety centrifuge cups, biological safety cabinets, etc.

Personal Protection Equipment (PPE): gloves, lab coats, goggles, face shields, etc.

BSCs type II: protect workers when used in combination with good microbiological techniques, & prevents external contamination by materials being manipulated inside cabinet

Biological Safety Cabinets (BSCs) type II

Safety Equipment

Maintain constant air curtain

Minimize movements in/out

No Burners inside the BSC

Disinfect working surface and interior

Avoid clutter, keep grille clean

UV lamps will be used with care and only for 30 min.

Do not use volatile chemicals inside the BSC

Certify performance annually (We annually certify our BSC at NMU)

Proper use of Biological Safety Cabinets (BSCs)

Secondary containment Protection of external environment

Facility Design

Contributes to the protection of people working in labs

Provides a barrier to protect persons outside the labs

Protects persons or animals in the community

Operational Practices

Determined by Risk Group of biohazard agents

RISK GROUP 1

Agents not associated with disease in healthy adult humans

Primary Barriers No safety equipment required

Secondary Barriers Open bench and sink

BioSafety Level 1 (BSL-1)

• Limited laboratory access unless responsible permits• No eating, drinking, applying makeup, etc.• No mouth pipetting• Safe handling of sharps• Safety glasses • Lab coats (must stay in the lab until washed)• Hand washing using antimicrobial soap• Decontaminate spills, cultures and waste

Good Microbiological Practices for BSL-1

CDC guidelines for BSL 1 labs“Access to the laboratory is limited or restricted at the discretion of the laboratory director when experiments or work with cultures or specimens is in progress.”

Handwashing with soap and water remains a sensible strategy for hand hygiene in non-health care settings Hand Hygiene Guidelines Fact Sheet , CDC

Hand washing for Safety

Twenty seconds of vigorous washing with soap and water

Method:

When:After lab section or as often as needed

Disinfection Practices

When?Before and after lab experimentTo decontaminate spills

How?Add disinfectant and wipe with a paper towel

In the case of a spill, apply disinfectant around the spilland wipe with the paper towel from the periphery to the center

Discard paper towels in the biohazard bag

Good Laboratory Practices of BSL-1 plus…

Limited access

Biohazard warning signs

'Sharps' precautions

Biosafety manual defining any needed

waste decontamination

RISK GROUP 2

Agents associated with human disease that is rarely serious or for which preventive or therapeutic interventions are often available

BioSafety Level 2 (BSL-2)

Into biohazard bags:• All cultures and biological fluids• All contaminated glass• All needles & syringes• All other potential biohazard material

Filled biohazard bags are autoclaved for 40 min. at 121oC & 15 lbs/in2

After biohazard bags and cardboards are autoclaved, they are bagged in black plastic bags

Procedures for disposal of Biohazards

Autoclave Guidelines

Equipment must continuously monitor & record temperature & pressure during entire length of each cycle.

Temperature-sensitive tape must be affixed to each bag or

container. Effectiveness must be evaluated under a full load at least once

every 40 hours of operation

Each bag must be exposed to a minimum of:– 121 degrees Celsius– 15 pounds of pressure per square inch– At least 30 minutes time AT THESE CONDITIONS

Autoclave Recordkeeping

A written log shall be maintained for each unit & shall contain the following:– Date, time, duration, & operator of each cycle– Approximate weight/volume of medical waste treated during

each cycle– Temperature & pressure maintained during each cycle– Method utilized for confirmation of temperature & pressure– Dates & results of calibration & maintenance

Written log record must be retained for three years

You always have the right to ask questions, or report hazards, either directly or anonymously without any fear of reprisal to…

Your lab instructor and/or the course instructor

Biology Department Head and Chemical Hygiene Officer – Dr. John Rebers (jrebers@nmu.edu)

Public Safety (227-2151)

Right to Know Policy

Next…

• Please complete the PRESENTATION REVIEW ACKNOWLEDGEMENT

• Then complete the QUIZ that follows the confirmation

• You must achieve a perfect score on the quiz to successfully complete your safety training. You may retake the quiz as many times as necessary.

• Failure to complete your safety training will result in your exclusion from the laboratory portion of your course

After the quiz…

• Remember to complete any additional Safety Training modules that are required for your course.