biological safety in...

Biological Safetyin laboratories

THE CHALLENGE

Safe Labs

• What is ‘safe’?

• Who decides?

• How to inform occupants?

• How to manage change?

•3

Involves & relies on

LAB/RESEARCH COMMUNITY SAFETY

•TeachingLaboratories

•LaboratoryResearch

•Research Field

•Chemical Hygiene Officer

•Radiation & Bio Safety Officers•Fire Prevention, Building Design &

Maintenance, Industrial Hygienists etc.

is responsible for quality and safety

EVERYONE

SAFETY, HEALTH & ENVIRONMENTAL GOALS

ZERO…• Workplace injuries• Workplace illnesses• Related injury/illness at home• Property loss• Unintended environmental damage

MANAGEMENT OF HEALTH, SAFETY AND THE ENVIRONMENT

Achieving excellence in providing a healthy and safe working environment

Supporting environmentally sound practices in the conduct of University activities

Comply with all applicable health, safety, and environmental protection laws, regulations and requirements

BIOLOGICAL RESEARCH IS ANY EXPERIMENTAL ACTIVITY

INVOLVING THE FOLLOWING BIOLOGICAL AGENTS:

• Human source materials (includes primary and immortal cell lines, clinical specimens, samples, tissues, etc)

• Animals and animal source materials• Plants and plant source materials• Recombinant and synthetic DNA/RNA (including all viral

vectors, RNAi technologies, etc).• Microorganisms (bacteria, viruses, fungi, prions, yeast,

algae, rickettsiae, parasites, etc)• Biological toxins, allergens

RG1 Agent not associated with disease in healthy adult humans e.g. Bacillus subtilis

RG2 Associated with human disease which is rarely serious and preventive or therapeutic interventions are often available e.g.Staphylococcus aureus, influenza viruses type A, B, and C

RG3 Serious or lethal human disease; preventive or therapeutic interventions may be available e.g. Mycobacterium tuberculosis

•RG4 Serious or lethal human disease; preventive or therapeutic interventions are usually not available e.g. Ebola

The CDC Biosafety in Microbiological and Biomedical Laboratories (BMBL) classifies risk group agents to:

Quick questionWhich risk group do you think the avian flu virus would belong to?

Resources for assigning risk group/biosafety level

1. The NIH Guidelines Appendix B assigns risk groups to some biological agents.

2. The American Biological Safety Association (ABSA) website provides a searchable database of biological agents and their assigned biosafety levels by country.

BIOSAFETY LEVELS

BL-1: Agents are not known to cause disease

BL-2: Agents are associated with human disease

BL-3: Agents are associated with human disease and are potentially transmitted as aerosols

BL-4: Agents of life threatening nature

BIOSAFETY LEVEL 1 (BL-1)

Use BL-1 when working with:• Well characterized agents• Agents that are not known to cause disease in

healthy humans • Agents that are of minimal hazard to lab

personnel and the environment

Examples of BL-1 Agents:E. coli JM109, DH5aSaccharomyces cerevisiae


Standard Work PracticesUse mechanical pipetting devicesWash hands frequentlyMinimize splashes and aerosolsDecontaminate work surfaces dailyHandle wastes properlyMaintain insect and rodent control

program

BIOSAFETY LEVEL 1 (BL-1)Personal Protective Equipment (PPE)Lab coat or apronSafety glasses or gogglesGloves as needed


Use BL-2 practices when working with:Agents of moderate potential hazard to personnel

and the environment

Examples of BL-2 agents:Human blood or body fluidsE. coli 0157:H7Clostridium botulinumRetroviral vectorsHuman cells in cell culture


RequirementsAdequate illuminationEyewash facilityNegative air pressureAutoclave availableBiological safety cabinet (BSC)Lab must be separated from public areas

BIOSAFETY LEVEL 2 (BL-2)Standard Work PracticesUse mechanical pipetting devicesWash hands frequentlyMinimize splashes and aerosolsDecontaminate work surfaces dailyHandle wastes properlyMaintain insect and rodent control

program

BIOSAFETY LEVEL 2 (BL-2)Special PracticesPlace used slides and coverslips in sharps

containers, never in any other receptacle.Sharps containers are:

• Red in color• Marked with the biohazard symbol• Puncture resistant• Leak proof

BIOSAFETY LEVEL 2 (BL-2)Personal Protective Equipment (PPE)• Lab coat or apron• Safety glasses or goggles• Gloves• Biosafety cabinet

CASE STUDY

CASE STUDYOpinion in favor of Dr Harran:1. Why should he be responsible? Most

professors don’t provide their graduate students much safety advice anyway.

2. Sangji should have worn a lab coat without the instruction of her advisor, it’s one of the basics of Organic chemistry and she is a graduate student not an undergraduate.

3. Sangji should have read the MSDS for tert butyl lithium, which states that she should not use a plastic syringeand should use a separate glass syringe for each transfer.

CASE STUDYOpinion against Dr Harran:1. Sangji did not have enough experience to

perform the experiment on her own.2. Dr Harran’s lab did not conform to safety codes.3. Dr Harran should have advised Sangji to read

the material safety data sheets for the chemicals she was using.

LESSONS LEARNED

Importance of safety audits and inspections by specialized personnel.

Changing the culture, i.e. The system should help enforce safety by empowering students rather than dictating it from on high.

Biosafety cabinets (BSC) control airbornecontaminants during work with infectiousmaterial through the use of laminar airflowand high efficiency particulate air (HEPA)filtration.

BIOSAFETY CABINETS (BSC)

It is designed to protect personnel and theenvironment from infectious materialsinside the cabinet and to protect thematerial inside the cabinet fromcontamination from the lab environment.



Provide product, personal and environmental protection.

Various classes of BSCs are available• Amount of air exhausted• Amount of employee protection

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BIOSAFETY CABINETS (BSCS)Sterile air is exhausted from BSCs through

a HEPA filter.• HEPA filter can trap particles to 0.3 microns• Chemical vapors and fumes pass through HEPA

filters

BSC LOCATION IN THE LABORATORYSince the air curtain created at the front of thecabinet can be easily disrupted, a BSC should belocated away from air supply registers, entrances,high traffic areas, and laboratory equipment, e.g.centrifuges, that create turbulence.

SAFE AND EFFECTIVE USE OF THE BSCBefore beginning work:a.Monitor alarms, pressure gauges, or flow indicators for any changes.b.Turn the cabinet on and let it run for 3-5 minutes.c.Wipe work surface with an appropriate disinfectant, e.g. 70% ethanol.d.Plan your work and place everything needed for the procedure inside the BSC.e.Wipe items with disinfectant before placing in BSC.

SAFE AND EFFECTIVE USE OF THE BSCAvoid airflow disruption that could affect the level of protection provided by the BSC:a. Keep the BSC free of clutter, e.g. extra equipment and

suppliesb. Don’t place objects over the front air intake grille.c. Don’t block the rear air intake grille.d. Limit traffic in the area when the BSC is in usee. Make sure lab door is closed, and avoid opening and

closing door if located near the BSC.f. Move arms slowly when removing or introducing

items.g. Keep all materials at least 4 inches inside the sash.h. Don’t operate a Bunsen burner in the cabinet.

SAFE AND EFFECTIVE USE OF THE BSC

While working:

a.Work as far to the back of the BSC workspace as possible.

b.Segregate contaminated and clean items. Work from “clean to dirty.”

c.Clean up all spills in the cabinet immediately. Allow cabinet to run for 3-5 minutes before resuming work.

SAFE AND EFFECTIVE USE OF THE BSC

After completing work:

a. Wipe down all items with an appropriate disinfectant before removing.

b. Remove all materials and wipe all interior surfaces with an appropriate disinfectant, e.g. 70% ethanol.

c. Periodically decontaminate under work grilles.

BIOSAFETY CABINETS (BSCS)

Helpful Hints Enter straight into the cabinet, avoid

sweeping motions Don’t place materials on the grill Keep discard pan or bag inside the

cabinet

CASE STUDY

Lessons learned:

1. Never add any water to an unmarked container.

2. Label everything with the common chemical name.

3. Always attach your waste label to the waste bottle before you add the waste and fill it out completely and accurately.

PROTECTIVE PERSONAL EQUIPMENT (PPE)

CONTAMINATION ROUTES

•Ocular invasion

•Inhalation

•Ingestion

•Skin penetration

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PERSONAL PROTECTIVE EQUIPMENT

The Strategy of Last ResortRespiratory ProtectionEye, Face, Hand, FootProtective ClothingHearing Head Protection

The Occupational Safety and Health Administration (OSHA)

requires that personal protective equipment (PPE) be selected, applies to work done with animals too

Eye/face/mucous membrane protectionWhen working with potentially infectious materials outside of a

biological safety cabinet mucous membranes must be protected by wearing a surgical-type with safety glasses

Foot/skin protection Open toed shoes, sandals and other open footwear is prohibited.

Shorts are forbidden in laboratories using potentially infectious materials.

Protective lab coats are to be worn while in the laboratory. This protective clothing is removed and left in the laboratory before leaving for non-laboratory areas (e.g., cafeterias, library and administrative offices.)

PERSONAL PROTECTIVE EQUIPMENTHand protection GLOVES ARE NEVER A SUBSTITUTE FOR THOROUGH HAND

WASHING WHEN ENTERING THE LAB, AFTER REMOVING GLOVES, AND BEFORE LEAVING THE LAB. WASH YOUR HANDS FOR A MINIMUM OF 30-60 SECONDS WITH COPIOUS AMOUNTS OF LIQUID SOAP AND WATER.

OTHER REMINDERS:- Decontaminate work surfaces daily and after a spill

involving any viable material.

- Properly autoclave your waste. Do not allow waste to accumulate.

PERSONAL PROTECTIVE EQUIPMENTBSL 1 and 2 laboratory coatglovesgoggles or face shieldmaskshearing protection

•Laboratory Safety Orientation, 2009 •41

GLOVE SELECTIONCHEMICAL COMPATIBILITY

http://www.ehs.okstate.edu/hazmat/gloves.htm

www.northsafety.comhttp://www.showabestglove.com/site/chemrest/www.ansell-edmont.com/download/Ansell_7thEditionChemicalResistanceGuide.pdf

QUESTIONIf you wear contact lenses in the laboratory:

A. take them out before starting the lab.

B. Keep them on but wear protective goggles.

C. advise your science instructor that you wear

contact lenses.

D. keep the information to yourself.

QUESTIONPersonal eyeglasses provide as much

protection asA. a face shield.

B. safety glasses.

C. splashproof chemical goggles.

D. none of the above.

WASTE DISPOSAL

BIOLOGICAL WASTE

• Liquid Waste• Sharps• Solid biohazardous waste

LIQUID WASTE

• Collect in containers with lids

• Autoclave or treat with 10% bleach to sterilize

• Dispose of down the drain (as long as no chemical or radioactive contaminants are present)

LIQUID WASTE• Procedure:• Collect BL1 and BL2 liquid waste in autoclavable, leak

proof containers that are never more than¾full.• Place containers in an autoclavable tray in the autoclave.

LOOSEN each container top and place indicator tape on each top.

• Adequate cycle time varies depending on load, type of autoclave, and secondary containment. Based on spore testing, determine the

• Appropriate cycle time to sterilize liquid waste for your autoclave. Typical cycle times for sterilizing liquid waste range from 45 to 90 minutes at 250°F.

• Autoclave temperature should be 250°F (121°C) and autoclave pressure should be 15 psi.

• Pour sterilized liquid waste down the sink and flush the drain with water.

SHARPSSharps (needles, syringes, scalpel blades,

slides, blood vials, pasteur pipettes)•Collect in approved sharps container

•Autoclave to sterilize

•Dispose of in medical waste boxes

SOLID BIOHAZARDOUS WASTE• Collect BL1 and BL2 waste in a red bag-lined Stericycle biowaste box or reusable bin for pick-up and off-site treatment. The inner red bag must be tied closed.

• If using cardboard boxes, close the box with tape and write your building and room number on the side of the box. If liquid is placed in the Stericycle boxes or reusable bins, then place enough absorbent material in the box to absorb the liquid to prevent leaking.

• Biowaste boxes or reusable bins are picked-up for off-site Electro Thermal Deactivation (ETD)

SAFE WORK PRACTICES

SAFE WORK PRACTICES

•WHAT SHOULD IDEALLY BE DONE

•WHAT WE CAN ACTUALLY DO

WORK PRACTICES

Education• Hazard recognition and control methods.

Training• Proper techniques; emergency response

Supervision- good safety performance

The biological safety program of the institution aims at:•Promoting a safe and environmentally-friendly research environment.

•Developing institutional and lab specific policies on the safe handling of biohazardous materials, biosafety levels, and personal protective equipment requirements.

•Providing training, risk assessment consultations, and laboratory safety audits.

•Assisting scientists in the adherence to all federal, state, and local requirements for biological research.

BIOLOGICAL SAFETY PROGRAM

PRIORITIES ARE:•Preventing laboratory acquired infections (LAIs)•Facilitating research compliance•Protecting the environment•Raising awareness on the importance of laboratory safety.

BIOLOGICAL SAFETY PROGRAM

GUIDING PRINCIPLES1. Management Commitment and Involvement of

Faculty, Staff, and Students2. Management Responsibility for Safety and the

Environment3. Establishing Clear Roles and Responsibilities4. Identification of Safety and Environmental Standards and Requirements5. Obtaining Authorization Prior to Conducting an Activity

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ADMINISTRATIVE RESPONSIBILITIES

Medical• Surveillance and immunization.

Hazard Identification• Signs - notifications, etc.

MEDICAL SURVEILLANCE

A medical surveillance program is provided throughUniversity Health Services (UHS) for personnelwho are occupationally at risk of exposure to bloodborne pathogens (BBP).

UHS provides medical treatment and post-exposure evaluation after an exposure to infectiousagents or rDNA for students and employeesworking in BL1 or BL2 laboratories.

e.g. free hepatitis B vaccine offer and post-exposure evaluation with follow up.


A medical surveillance program is also available forpersonnel that have direct contact with researchanimals

The program includes disease-specificoccupational health advice, Animal Biosafety Level2 training, vaccination offers, pre-screeningquestionnaires, and post-exposure medicaltreatment.


In addition to being offered recommendedvaccinations, BL3 laboratory personnel may havebaseline serum samples collected, as appropriate,and tests for agents handled in the lab, e.g. TB skintest.

•NOT APPLICABLE IN OUR INSTITUTION


SAFETY AUDITS9 INSPECTIONS

Labs inspected for compliance with • OSHA Lab Standard

• Chemical Hygiene Plan

• Hazardous, Medical and Radioactive Waste• Labeling, Storage & Segregation

• Radioactive isotope use• Biohazardous materials use• Electrical Code

SAFETY AUDITS9 INSPECTIONS The Harvad Biosafety office must:Conduct lab inspectionsDevelop emergency and reporting

proceduresInvestigate lab accidentsReport incidents and violations of the

guidelines.Provide biosafety training.

LABORATORY BIOSAFETY AUDITS SUGGESTED IN OUR INSTITUTION

Assessing appropriate personal protective equipment

Proper use of laboratory equipment (biological safety cabinets, centrifuge, etc)

Proper labeling of equipment for the storing/manipulation of biological materials

Proper procedures for decontamination of biological waste (solid, liquid, sharps)

Presence of life safety devices (eyewash, safety shower, antimicrobial soap, etc)

AUDIT PROCESSContact department and/or lab representative

• Schedule audits for that dept/labOccupants present

• Yes – can ask more – takes longer – better result• No – limited to physical conditions only

Preparation of reports

5 Steps of Integrated Safety and Environment Management System (ISEM)(CORE FUNCTIONS)

1. Define scope of activity

2. Identify & analyze hazards

3. Develop & implement controls

4. Perform activity within controls

5. Provide feedback & make improvements

QUESTIONYour laboratory specializes in virology. There is an epidemic of H5N1 in your country and the Ministry of Health names your laboratory as the reference laboratory for processing all samples. What are the elements to consider ensuring biosafety in your laboratory?

EMERGENCY PREPAREDNESS


•Individual preparedness

•Common hazards

•Emergency procedures

•Emergency communications

•Emergency resources


The primary areas of focus include:

•Inclement weather

•Natural disasters

•Chemical or biohazardous disasters

•Fires and floods

•Suspicious behavior/activity

WHAT TO DO IN THE EVENT OF AN INJURY OR POTENTIAL EXPOSURE:

HELPFUL RESOURCESThe Biosafety in Microbiological and Biomedical Laboratories

(BMBL) is a CDC/NIH publication that became the cornerstone of biosafety practices and policies in the United States

The NIH Guidelines for Research Involving Recombinant or Synthetic DNA

The OSHA Bloodborne Pathogen StandardWHO Biological Safety Manual

biological safety in...

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