Mayfield_BSL2_Manual_2023_Nov_MostCurrent

General Laboratory Practices for BSL-2 Laboratories

1. Dayne Mayfield Lab

A general template for this document can be found here.  The principal investigator is responsible for including laboratory and protocol specific procedures for addressing hazards in their laboratory.  This plan must be adopted by the Institutional Biosafety Committee (IBC).

The official “University of Texas Laboratory Safety Manual” is located on the top shelf above the desk in MBB 1.124.  Please refer to it if you require a more in-depth description of university safety responsibilities, emergency procedures, fundamentals of laboratory safety, and basic and/or specific procedures for working with chemicals and biological hazards

Additionally, the EHS website is has the most up-to-date information. Always consult it if you have any questions/concerns.

This BSL2 manual must be reviewed and signed annually by all lab members.

I have read, understand, and agree to adhere to the biosafety procedures contained within:

Printed Name              Signature                     Date

PI:

_Mayfield, R. Dayne_               

Written (Date):  _______________

Last Revision (Date):  __2/23/23__

Reviewed by BSO (Date):  ____________

Adopted by the IBC (Date10/19/2022): 

Table of Contents

Principal Investigator Responsibilities………………………………………...…    3

Laboratory Staff/Student Responsibilities…………………………………….....    4

General Emergency Information……………………………………….………...    4

Emergency Contacts……………………………………….…………………….     4

Fire Alarms/Extinguishers……………………………...……….………………..    5

Eyewashes……………………………………….……………………………….    5

Spills in the Laboratory……………………………………….………………….    5

Biological Spill……………………………………….…………………………..    6

Spills Outside the Biosafety Cabinet……………………………………….…….    6

Spills Inside the Biosafety Cabinet…………………………………………..…..    6

Spills Inside Equipment……………………………………….………………….   6

Chemical Spill……………………………………….……………………………   6

Radiation Spill ……………………………………….………………...…………   7

Exposures in the Laboratory……………………………………….…………….     7

      Biological Exposure……………………………………….……………………..    7

Chemical Exposure……………………………………….……………………...     7

      Radiation Exposure……………………………………….……………………...    8

BSL-2 Safety Procedures……………………………………….………………..    8

New Employees……………………………………….…………………………     9

Training……………………………………….………………………………….    9

Medical Surveillance……………………………….…………………………….    11

PPE……………………………………….………………………………………    11

Biohazard Warning Signs and Posting……………………………………….…..    12

Biological Safety Cabinets (BSC’s) ……………………………………….…….    12

Biological Waste Disposal……………………………………….………………    13

Chemical Safety……………………………………………………………….…    13

Fume Hood Use……………………………………….…………………………     14

Housekeeping………………………………………………………………….…    14

Laboratory Close Outs/Equipment Disposal…………………………………….     14

Security……………………………………….………………………………….     14

Controlled substances……………………………………………………………     15

Select Agents/Toxins………………………………………………………….…     16

Sharps……………………………………….……………………………………    16

Shipping Biological Material……………………………………….……………    17

Transporting Biological Material……………………………………….………..    17

ClusterMarket…………………………………………………………………….    17

Useful Web Sites……………………………………….………………………...    17

Appendix A……………………………………….…………..………………….    18

Appendix B……………………………………….………………………………    20

Appendix C……………………………………….………………………………    21

Principal Investigator Responsibilities

The Principal Investigator (PI) has the primary responsibility for ensuring that their laboratory is safe.  They must adhere to all guidelines and regulations.  They are responsible for the safe use of biological, chemical and radioactive materials in their laboratory.

In addition, the PI must:

            Limit personnel, student, and visitor exposure to hazards to the lowest practical level.

            Provide special safety considerations for individuals under the age of 18

            Apply the recommended biosafety level for the work being conducted

            Be familiar with the required medical surveillance for each type of agent in their laboratory and ensure staff/students/visitors have medical clearance

            Develop written lab specific safety procedures and train their personnel on them 

            Maintain documentation of training

            Provide PPE and instruction on use

            Ensure waste is properly disposed

            Report spills, exposures or incidents to Environmental Health & Safety (EHS)

            Conduct periodic drills of emergency procedures

Laboratory Staff/Student Responsibilities 

            Know the biological materials and procedures used in the laboratory

            Follow approved lab procedures and safety guidelines

            Know emergency procedures

            Complete all required training before conducting any lab activity

            Complete recurring trainings by the due-date

            Report any unsafe conditions to the PI, EHS or the RRT

            Utilize all required Personal Protective Equipment (PPE)

            Use appropriate lab equipment and containment facilities

General Emergency Information

Emergency Contacts

If there is an emergency, call 911 to reach UTPD.  UTPD will then summon assistance from the City of Austin Emergency Services.  If there is a fire or explosion that you cannot control, evacuate the area immediately.  Pull the fire alarm and then call 911 from a safe location.  For other emergencies, dial 911 for UTPD.

If any emergency or significant spill/exposure occurs in the laboratory, immediately notify EHS and your lab supervisor/PI. ALL lab incidents (spills, injuries, near-misses) should be reported to EHS using their online form. Phone support (24hr) is also available at 512-471-3511.

Fire Alarms/Extinguishers

Locations of fire pull station alarms:

(1)  Exterior entrance – south wing of first floor

(2)  South end of first floor hallway

(3)  Loading dock exterior door

Location of fire extinguishers:

(1)  MBB 1.124 next to telephone

(2)  MBB 1.124 next to door for 1.124B

(3)  Hallway

(4)  MBB 1.106 next to the door

Know the location of each of these, and identify the location of the extinguisher closest to your lab bench.  If the fire alarm sounds, leave the building immediately and move away to a safe distance.

Eyewashes

Location of eyewashes: MBB 1.106 sink, MBB 1.24 sink and MBB 1.128 (cold room) sink

In case of exposure, proceed to nearest eyewash station.  Hold eyelids open with thumb and forefinger and rinse for at least 15 minutes.  Wash from the outside edges towards the inside to prevent washing chemicals back into the eye. 

Rinse should be aimed at the inner corner of the eye (near the nose) not directly at the eyeball.  “Roll” eyes around and up and down to ensure full rinsing. 

Contact lenses (if worn) should be removed as soon as possible.  Have another member of the lab call for emergency response immediately.  The area around the eye wash station must remain clear at all times.

Spills in the Laboratory

Call EHS when a significant spill occurs.  A lab incident report form (EHS) must be filled out for significant spills. 

A significant spill is defined as:

• Spills greater than 5 mL (BSL-2, toxic chemicals) outside primary containment

• Spills that result in an exposure

• Spills that present an inhalation hazard

• Spills that cannot be easily cleaned

• Spills that endanger people or the environment

Biological Spills

Location of universal spill kits: MBB 1.124 [shelf over freezer #3], MBB 1.118 [on top of flammable cabinet] and MBB 1.106 [under the sink]

Notify your laboratory supervisor/PI and nearby lab staff.  Use appropriate PPE when cleaning.  Dispose of all cleanup material as biohazardous waste. 

Spills Outside the Biosafety Cabinet

Decontaminate biological spill by covering the spill with paper towels and soaking in a fresh 10% bleach solution or disinfectant for at least 20 minutes. 

Clean area at least 2 feet around spill with disinfectant.  (Including furniture/walls)

Spills Inside the Biosafety Cabinet

Decontaminate any biological spill by covering the spill with paper towels and soaking in disinfectant for at least 20 minutes. 

While spill is soaking, wipe down all work surfaces and equipment in BSC with disinfectant.  Place all cleanup material into biohazardous waste container.

Spills Inside Equipment/Instruments

If the spill occurs in a shaker or centrifuge, turn off the equipment and leave the door closed for at least 30 minutes to reduce aerosol exposure before cleaning up the spill.  Tape a sign indicating the problem and your name to the contaminated equipment until the situation has been rectified. 

Chemical Spills

Location of spill kit:   MBB 1.124 [shelf over freezer #3]

Clean up all chemical spills immediately.

If you spill a hazardous material and need assistance, call EHS. 

Radiation Spills [Note: we are not currently using any radioisotopes]

Location of spill kit (LiftAway):  MBB 1.136

Wipe up the spill with Lift-away and check for residual contamination by performing a wipe test (tritium or carbon-14) of the affected area.  Monitor with a portable Geiger counter capable of detecting the energy signature of the radioisotope. 

For other radioisotopes, refer to the EHS Radiation Safety Manual for proper detection methods. 

Call EHS if you need assistance.

Exposures in the Laboratory 

All exposures must be reported to the PI and to EHS. Contact OHP. 

Biological Exposure

Remove any contaminated clothing/jewelry and wash skin exposed to the agent with a disinfectant such as antibacterial soap. 

Autoclave any contaminated clothing before disposal.  Decontaminate any surfaces using the procedure for a biohazardous spill. 

If you believe you may have been exposed to an agent, contact EHS and seek immediate medical attention.

If you suspect you have a lab-acquired illness regardless of a recent exposure, see a health care professional immediately.  Provide the health care professional with EHS contact information to allow for coordination with the university safety office. 

Fill out a worker’s compensation form even if you are not sure if your illness was acquired at work.  This must be done if you need to file a claim later.

Chemical Exposure

For most chemical exposures to the skin, wash the chemical off with water for at least 15 minutes and remove any contaminated clothing.  Use the sink for small exposures; for large exposures use the safety shower. If using the safety shower, call EHS immediately; they have supplies to help mitigate flooding.

Location of safety shower:  Hallway next to MBB 1.122

Radiation Exposure

For skin contamination with radioactivity, wash it off and remove any contaminated clothing.  If you believe you have inhaled or ingested any radioactivity, call EHS.

BSL-2 Safety Procedures

This lab is rated as Biological Safety Level 2.  BSL2 is required for work involving agents of moderate potential risk to personnel and the environment. 

Eating, drinking and smoking are prohibited in BSL-2 laboratories.  For additional requirements consult Biosafety in Microbiological and Biomedical Laboratories, 6th Edition. The CDC also has some video training that may be useful.

Types of Biohazards or Potentially Infectious Materials:

1.  

   1. Human, animal and plant pathogens:

• Bacteria, including those with drug resistance plasmids

• Rickettsiae

• Fungi

• Viruses, including oncogenic viruses and viroids

• Parasites

• Prions

1.  

   1. All human and/or non-human primate blood, blood products, tissues and certain body fluids.

   2. Cultured cells (all human and certain animal species) and potentially infectious agents these cells may contain.

   3. Biological toxins (bacterial, fungal, botanical, etc.)

   4. Certain recombinant products

   5. Infected animals and animal tissues

Biohazardous Materials:  Provide a description of material used in the laboratory in Appendix A.  Include symptoms/hazards/medical surveillance for working with the material.

Potentially Hazardous Equipment: Provide a description of potentially hazardous equipment used in the laboratory in Appendix B.  Examples include sonicators, blenders, lyophilizers, Bunsen burners, etc…

Special Practices: Provide a description of potentially hazardous procedures conducted in the laboratory in Appendix C.  Examples include culture extraction, centrifugation, use of sharps, etc…

New Employees

Location of lab manual:  MBB 1.124, above the desk; in the Lab Safety and Training folder on Box

Laboratory personnel should be aware of the potential hazards associated with the work and be proficient in the specified practices and procedures.

Know the chemicals you are working with. Look up the material safety data sheet (MSDS) available at https://ehs.utexas.edu/programs/labsafety/sds-chemical-information.php. Additional MSDSs can be obtained directly from the specific vendor supplying the reagents. Place a bookmark on your computer so that you can access this information quickly. The EHS website has a list of common non-hazardous chemicals and a searchable chemical compatibility chart can be found here.

For biological agents, https://www.canada.ca/en/public-health/services/laboratory-biosafety-biosecurity/pathogen-safety-data-sheets-risk-assessment.html is an easily searchable site from the Canadian government. Also see the NIH Guidelines for Recombinant or Synthetic Nucleic Acids. Information from the Institutional Biosafety Committee ishere.

If you are using an instrument/equipment for the first time, please ask for instructions/training. Immediately report broken or malfunctioning instruments/equipment to the lab manager.

Training

All laboratory research personnel must take institutional provided training. Training must be documented (electronic or paper).  Personnel should not initiate research until training is completed. All classes are accessed online via UTLearn, except where noted.

Initial Training for all Lab Personnel: