Potentially Hazardous Biological Agents

MOST IMPORTANT: You MUST get formal PreApproval by our SRC before you may begin handling PHBAs. Florida does NOT recognize SRC exemptions for cultured microorganisms.

• Our Scientific Review Committee (SRC) reviews proposed research projects for safety and ethics. They consider the researchers' safety, containment and disinfection, and public health needs for research done on PHBAs.

• PHBA projects (even food molding) may NOT be conducted at home.

• PHBA projects must be assigned a BioSafety Level (BSL).

  • Middle school students are only allowed to conduct BSL-1 projects. High school students may conduct BSL-1 or BSL-2 projects.

  • Science classrooms are appropriate for BSL-1 projects. But BSL-2 projects require a BSL-2 rated facility.

    • Please note: Only one SCPS school has a BSL-2 rated facility; almost all BSL-2 projects must be conducted at a professional lab environment, which requires permission from a professional willing to oversee the project (a Qualified Scientist) AND someone in management / an administrator at the facility. Some technologies also necessitate that the facility is part of a regulated research institute such as UCF or Nemours.

  • The overall BSL of your project is the HIGHER of: the BSL of the species cultured and the BSL of the procedures used.

    • For precollege student research, to stay BSL-1, the student may NEVER open a culture after inoculation. The culture must be sealed (taped) just after inoculation and NOT reopened. At the end of experimentation, the teacher/supervisor may untape the culture and place it in the bleach bath for disinfection then dispose of it.

• IMPORTANT: Almost every single project will need at least one round of revisions before the SRC approves it. Use the info below to pre-empt the most common issues! But please factor this into your timeline when planning your project. 

The deadline to submit your project to our SRC for project PreApproval is 10/25/2024. Exception: AP Research students have a separate deadline of 11/22/24 to follow the CollegeBoard course timeline. Earlier submission is STRONGLY encouraged.

BSL-1 Requirements

• If your project is looking at MOLD GROWTH on food items: 

  • This is the one kind of PHBA project that doesn't have to come through our SRC for PreApproval. As long as you follow the requirements below, you can do it without formal PreApproval.

  • This project must be done in a BSL-1 rated facility (so a science classroom is okay but NOT AT HOME).

  • Design the experiment so that you throw away each sample as soon as it develops mold.

    • You may NOT measure how moldy all samples are the end of a set time period.

    • You MAY measure how long it took for each sample to develop mold. But you must dispose of each sample as soon as that sample shows mold.

• If your project is looking at MICROORGANISMS and/or RECOMBINANT DNA:

  • Reminder: The project MUST be reviewed and approved by our SRC BEFORE you begin. Florida does NOT recognize exemptions when cultured.

    • IMPORTANT: Even though the ISEF rulebook (pg 17) lists E. coli K12, archaea, and protists as exempt, Florida's rules override that when cultured.

  • This project must be done in a BSL-1 rated facility (so a science classroom is okay but NOT AT HOME).

  • Plan that you will need to disinfect work surfaces with 10% bleach (or ethanol, if your lab prefers that) before and after use each day. Plan that you will need to wash your hands with soap and water before and after working in the lab each day.

  • Plan that you will need proper PPE (lab coat/apron, safety goggles/glasses, and nitrile/latex gloves). These must appear in the materials list, and you should have a procedure step saying when you'll put them on.

  • Plan that once you inoculate your culture into the container, you will seal it (tape) and NOT REOPEN IT.

    • Petri dishes have clear lids for a reason! At the end of your experiment, you'll need to count your colonies, estimate percent cover, or measure colonies/margins while looking through the lid (still sealed).

    • If using liquid cultures, that means you may need to run the whole experiment in spectrophotometer cuvettes. Plan accordingly!

    • If testing bactericidal properties of different chemicals, look into the Kirby-Bauer method. This is a BSL-1 compatible method where you soak little discs in the chemical and place them into the Petri dish during (or just before) inoculation.

  • Disposal: Your lab supervisor/teacher (not you) will remove the tape from the container at the end and place it into the 10% bleach bath. If the lid is screwed on, they should unscrew it while submerged. Everything should soak in the 10% bleach bath for at least 30 minutes, then solids should be thrown away in the trash, and liquids (including the used bleach bath solution) should be rinsed down the sink with lots of water.

    • Tools that you use along the way should also be soaked in a 10% bleach bath, on the day they were used, for 30 minutes then disposed.

    • Used glassware or other non-disposables can be soaked in the 10% bleach bath for 30 minutes then washed with soap and water for future reuse.

    • If you are doing your project at a professional lab, and they use an autoclave for disinfection, that's fine. Just don't autoclave bleach! Our autoclave requirements are 121 deg C at 15 PSI for at least 20 minutes.

• Overall note: Once you have a general idea of what you're going to do and how you're planning to do it, double check it against the ISEF AND SSEF rules. Go through the PHBA rules sections and re-read them, specifically making sure that your project doesn't violate any of them.

BSL-2 Requirements

• Only high school students (NOT middle school students) are allowed to conduct BSL-2 projects.

• In Florida's precollege research rules, it is considered an automatic BSL-2 procedure if the student researcher opens a culture after inoculation for any reason.

• BSL-2 projects MUST be done in a BSL-2 rated facility. Some project topics require that this is specifically within a regulated research institute. Locally, only UCF and Nemours are RRIs for this purpose.

  • NOTE: A BSL-2 lab must be restricted-access. That means it CANNOT be used as a teaching classroom while your project is in there.

  • Unless your high school has a BSL-2 facility, you MUST get permission to do your project at BSL-2 lab from BOTH the professional who agrees to supervise your project (your Qualified Scientist) AND an administrator / someone in management at the facility. The administrator may NOT be otherwise related to your project, and they will document this permission on BOTH the permission letter/email (attached to Form 1A) AND by signing as the administrator on your BSL-2 checklist.

• BSL-2 projects MUST be overseen by a Qualified Scientist (QS).

  • This is someone who has an advanced degree in a microbiology-related field (a Master's or Doctoral degree).

    • Science education degrees are not acceptable for a QS. However, if the school has a BSL-2 rated facility, then a non-QS science teacher with experience in this area can still serve as Designated Supervisor as long as a QS is willing to advise on the biosafety requirements and protocols for the project.

  • BEFORE you submit for SRC PreApproval, the QS will sign your Form 6A, BSL Checklist, and Form 3. They will also complete a Form 2. AFTER experimentation, they will complete and sign a Form 1C detailing what of the work/thinking was theirs vs yours.

• Project design:

  • Plan that you will need to disinfect work surfaces with 10% bleach (or ethanol, if your lab prefers that) before and after use each day. Plan that you will need to wash your hands with soap and water before and after working in the lab each day.

  • Plan that you will need proper PPE (lab coat/apron, safety goggles/glasses, and nitrile/latex gloves). These must appear in the materials list, and you should have a procedure step saying when you'll put them on.

  • If your project requires the use of a Biosafety Cabinet, be sure to list that in the materials list (including its brand and model #, or at least its Class #). 

  • Plan that tools that come into contact with PHBAs or PHBA-contaminated materials, they will be autoclaved at 121 deg C and 15 PSI for 20 minutes after use. After autoclaving, disposables should be placed in a sealed biohazard waste bag in the trash, and non-disposables should be washed and dried for future reuse.

• Disposal: At the end, cultures and tools should be autoclaved at 121 deg C and 15 PSI for 20 minutes. Then, for solids, disposables should be placed in a sealed biohazard waste bag in the trash, and non-disposables should be washed and dried for future reuse. After autoclaving, liquids may be able to be rinsed down the sink depending on what liquids were used.

  • You should NEVER autoclave bleach! You should also check the flash point for any chemicals that you think may be highly flammable.

    • Autoclaving, when safe, is the preferred disposal method for BSL-2 projects. If autoclaving is not safe, you may disinfect those particular samples by soaking in a 10% bleach bath for 30 minutes before disposing.

  • If you are working in a professional lab, and they use a biohazard waste pickup service, that's fine. You may still need to plan to autoclave tools / non-disposables. Just work with your QS to properly describe disinfection and disposal.

Documentation for PHBA Projects

• Research Plan 

  • You'll answer the subject-specific items for PHBA research. See the sections above to help with this, and READ the notes in the question prompts!

  • Your materials list should ALSO include your PPE and your disinfectant(s). Usually 10% bleach is used to disinfect work surfaces and for final container/tools disinfection before disposal. Some labs use ethanol for work surfaces instead. BSL-2 projects must use an autoclave instead for final container/tools disinfection before disposal. [If your BSL-1 lab has an autoclave, it's fine to use it.] Occasionally, professional labs contract out final disposal instead.

  • <Your procedure should ALSO include the steps for disinfecting work surfaces (10% bleach or ethanol), washing hands, and putting on PPE at the beginning. There should ALSO be procedure steps for disinfecting work surfaces and washing hands again at the end of each day. Your procedure should ALSO have the step(s) for  final container/tools disinfection and disposal at the end.

  • Your bibliography MUST include an applicable source about biosafety, chosen from the list on ISEF Rulebook. [The BMBL is the best.] Make this source #3 in your bibliography (after the ISEF rules and SSEF rules) so our reviewers are sure to find it!

• Form 2: Only required for BSL-2 projects.

  • For BSL-2 projects, this form must be completed BEFORE experimentation begins. It must be included when submitting your project for PreApproval review.

  • The Qualified Scientist is someone who has an advanced degree (Master's or Doctoral degree) in a microbiology-related field, who has agreed to supervise (either physically or just to advise on) the project.

    • If a different person will be the one physically overseeing your work in the BSL-2 lab, they are considered the Designated Supervisor. For PHBA projects, the Designated Supervisor must also have experience/training related to microbiological work.

  • The Qualified Scientist will fill out the top section, answer questions 1-4, and sign the bottom-left box.

    • If a different person will be the one overseeing your work (your science teacher, a grad student or post-doc, etc.), they should check No for #4. Then list the qualifications of that person (your DS). That person would then ALSO sign the bottom-right box.

    • Overall, if the professor / PI is actually supervising your work, some areas of this form will be blank. If they have someone else actually supervising your work, the whole form will be filled out (signed by both the professor/PI at left and the other person [DS] at right).

• Form 3

  • This will be repetitive, sorry. Summarize that culturing PHBAs always carries risk to the researcher, the final BioSafety Level, your precautions (PPE, washing hands before/after, disinfecting work surfaces before/after, disinfection before final disposal, etc), and your disposal method. Cite your biosafety source again (MUST match the one in your Research Plan Bibliography, so probably the BMBL). 

  • Mostly, this form will focus on chemical risks. If you are using a lot of different hazardous chemicals, you may need to use more than one copy of Form 3. If so, make sure that a particular risk's description, precautions, disposal, and source(s) appear on the same page.

  • If you have a Qualified Scientist (so BSL-2 projects), they must be the one to sign at the bottom of this form. Otherwise, have the person who will actually be supervising your work (Designated Supervisor) sign here. The person signing should still have experience/training in microbiological work.

• Form 6A

  • Summarize the information into questions 1-5! The text MUST be legible--if you use too many words, it will either cut some off or shrink it too small.

    • #1:

      • Purchased cultures: list species, source (company and item #), and BSL for each species purchased

      • Environmental swabbing: state "unknown microorganisms from environmental swabs of ________"

    • #2:

      • Science classroom (BSL-1) with sink for handwashing and lab-grade table surfaces

      • University research lab with restricted access and BSL-2 containment

    • #3:

      • PPE (list here), wash hands/disinfect surfaces before/after

      • If using a Biosafety Cabinet, also include that in the list as "Class __ Biosafety Cabinet"

    • #4: List as either BSL-1 or BSL-2

    • #5:

      • [BSL-1] Supervisor will remove tape and soak in 10% bleach for 30 minutes then solids in trash and liquids down sink

      • [BSL-2] Autoclave at 121 deg C and 15 PSI for 20 minutes then dispose in sealed biohazard bag in trash

  • For Section 2: Training, summarize info for questions 1-2.

    • Training #1: Standard microbiological practices including aseptic technique, chemical safety

    • Training #2:

      • BSL-1 example: "__ years certified science teacher"

      • BSL-2 example: "Ph.D. in ____________, __ years experience professional/university microbiology research"

  • For section 3:

    • If you don't know the correct answer to the check boxes, you can leave it blank, and our SRC will fill it in (unless it's the second check-box, but that's really rare, and your QS would know).

    • IMPORTANT: Have this part signed by the most-trained person overseeing your project.

      • If you have a professional mentor, they sign here.

      • If you have a professional mentor, and they have a grad student or post-doc who is actually with you while you're working, make sure the actual professor / PI is the one who signs here (Form 6A, section 3)

  • Section 4: Leave this blank, it's where our SRC's PHBA specialist signs!

• Form 6B: Most PHBA projects do NOT need this! You only need this if your project includes vertebrate animal tissue, cell lines, or body fluids (like blood). If you need this, have your research mentor fill out answers to 1-3, check the appropriate verify/certify box, and sign. 

• BSL Checklist: All PHBA projects DO need this.

  • MAKE SURE YOU USE THE CHECKLIST FOR YOUR FINAL BIOSAFETY LEVEL!

    • BSL-1 Checklist

    • BSL-2 Checklist

  • If you're doing the project at your school: Your teacher will fill this out and sign. A school administrator (usually the AP over science will also sign). All boxes should be checked Yes in order to qualify, but all SCPS science classrooms should meet these criteria.

  • If you're doing the project at a university lab or a professional lab: Your professional mentor at the lab will fill this out and sign. An administrator (someone in management) must also sign--that person MUST be the SAME person who wrote the permission letter (Form 1A attachment) stating you're allowed to do your project there. All boxes should be checked Yes in order to qualify [microbio labs at regulated research institutes like UCF and Nemours should all meet these criteria].

    • The facility name and address on the checklist should match what you listed at the bottom of Form 1A.

• AFTER FORMS: If you had a professional mentor and/or worked in a professional facility (so all BSL-2), that professional will need to fill out and sign Form 1C.

  • This should be dated AFTER experimentation. If you are still collecting data when you submit your project to apply to the fair, just have it dated near the submission date. NEVER forward-date (that could create version-confusion later on).

  • If they provided you remote advisement, please hand-write that at the top of the first page (or on the Student Name line).

  • If all you did was use facilities, or if all they did was advise your teacher on biosafety requirements, they should answer NO to question 1 and state that in 1a. Then they'd skip questions 2-5 and fill out the bottom box, sign, and date.

• Check out:

  • this BSL-1 sample paperwork file (annotated with instructions) and BSL-1 sample project file.

  • this BSL-2 sample paperwork file (annotated with instructions) and BSL-2 sample project file.