Students are permitted to do research projects with potentially hazardous biological agents meeting the conditions and rules described below which were designed to protect students and to ensure adherence to federal and international biosafety regulations and guidelines.

Index

Prohibited Studies
Rules
Projects Involving Unknown Microorganisms
Projects Involving Recombinant DNA (RDNA) Technologies
Documentation and Approval
Exempt Studies

Prohibited Studies:

1. Experimentation involving the culturing of potentially hazardous biological agents, even BSL-1 organisms and C. elegans, is prohibited in a home environment.
   1. However, specimens may be collected at home as long as they are immediately transported to a laboratory with the BSL containment determined by the affiliated fair SRC.
2. Students are prohibited from designing or participating in any research involving biosafety levels above BSL-2. (This includes BSL-2+, BSL-3 and BSL-4.)
3. Any study involving the collection and examination of body fluids that may contain biological agents belonging to a biosafety level over 2 is prohibited. (Please see Tissue & Body Fluid Rules)
4. Students are prohibited from the insertion of antibiotic- resistance traits or selection of organisms expressing traits that may affect the ability to provide effective treatment of infections acquired by humans, animals, or plants.
   1. Students are prohibited from designing or selecting for multiple drug resistant organisms (MDROs) to investigate the pathology, development, or treatment of antibiotic- resistant infections.
5. All studies involving the use of prions or prion-like proteins are This includes studies working with amyloid (A), tau, a-synuclein, transactive response DNA-binding protein of 43 kDa, and amyloid fibrils.
6. Propagation of recombinants containing DNA coding for human, plant or animal toxins (including viruses) is
7. Introduction or disposal of non-native, genetically- altered, and/or invasive species (e.g. insects or other invertebrates, plants, vertebrates), pathogens, toxic chemicals or foreign substances into the environment is prohibited. Students and adult sponsors should reference their local, state and national regulations and quarantine lists.

Rules:

1. Prior review and approval is required for the use of potentially hazardous microorganisms (including bacteria, viruses, viroids, rickettsia, fungi, cyanobacteria, and parasites) and recombinant DNA (rDNA) technologies.
2. Research determined to be at Biosafety Level 1 (BSL- 1) must be conducted in a BSL-1 or higher laboratory. The research must be supervised by a trained Direct Supervisor or a Qualified Scientist. The student must be properly trained in standard microbiological practices.
3. Research determined to be a Biosafety Level 2 (BSL- 2) must be conducted in a laboratory rated BSL-2 or above and follow BSL-2 safety conditions throughout the study. (Commonly limited to a Regulated Research Institution (RRI). The research must be reviewed and approved by the Institutional Biosafety Committee (IBC) if the RRI requires the review. For a high school BSL-2 laboratory, the SRC must review and approve. The research must be supervised by a Qualified Scientist.
4. Laboratory studies involving the culturing of clinically significant multidrug resistant organisms (MDROs) must have a written justification for usage and be conducted at an RRI laboratory with a minimum of BSL-2 containment and documented IBC review and approval.
   1. Representative examples include, but are not limited to the following known agents: MRSA (Methicillin-Resistant Staphylococcus aureus), VISA/VRSA (Vancomycin Intermediate or Resistant Staphylococcus aureus), VRE (Vancomycin- Resistant Enterococci), CRE (Carbapenem Resistant Enterobacteriacae), ESBLs (Extended Spectrum Beta-Lactamase producing gram negative organisms), and fungi (yeasts or molds) with known resistance to antifungal agents.
   2. Extreme caution must be exercised when selecting and sub-culturing antibiotic-resistant organisms. Studies using such organisms, including BSL-1 organisms that may have originally been exempt from prior SRC approval, require at least BSL-2 containment.
   3. Insertion of antibiotic resistance markers for the clonal selection of bioengineered organisms is permitted, with the exceptions outlined in prohibited studies item #4.
5. The culturing of human or animal waste, including sewage sludge, is considered a BSL-2 study.
6. Naturally-occurring plant pathogens may be studied (not cultured) at home, but may not be introduced into a home/ garden environment.
7. Projects involving water samples collected from active Harmful Algal Blooms are considered BSL-2 studies.
8. Insect and arthropod vector-borne pathogens such as Malaria, Lyme, are considered BSL-2 studies.
9. All local, state and national laws and permit requirements must be followed regarding the transport and use of microorganisms such as, but not limited to citrus greening or tobacco mossaic, etc.
10. All potentially hazardous biological agents must be properly disposed of at the end of experimentation in accordance with their biosafety level. For BSL 1 or BSL 2 organisms: Autoclave at 121 degrees Celsius for 20 minutes, use of a 10% bleach solution (1:10 dilution of domestic bleach), incineration, alkaline hydrolysis, biosafety pick-up and other manufacturer recommendations are acceptable.

Projects Involving Unknown Microorganisms:

Studies involving unknown microorganisms must adhere to the following rules:

1. Research with unknown microorganisms can be treated as a BSL-1 study under the following conditions:
   1. Organism is cultured in a plastic petri dish (or other standard sterile non-breakable container) and sealed.
   2. Experiment involves only procedures in which the petri dish remains sealed throughout the experiment (e.g., counting presence of organisms or colonies).
   3. The sealed petri dish is disposed of via autoclaving or disinfection under the supervision of the Direct Supervisor.
2. If a culture container with unknown microorganisms is opened for any purpose, (except for disinfection/ disposal), it must be treated as a BSL-2 study and involve BSL-2 laboratory precautions.

Projects Involving Recombinant DNA (RDNA) Technologies:

1. All rDNA technology studies involving BSL-1 organisms and BSL-1 host vector systems, including commercially available kits, must be conducted in at least a BSL-1 laboratory under the supervision of a Qualified Scientist or Direct Supervisor and must be approved by the SRC prior to experimentation. Examples include cloning of DNA in E. coli K–12, S. cerevesiae, and B. subtilis host-vector systems.
   1. An rDNA technology study using BSL-1 agents that may convert to BSL-2 agents during the course of experimentation must be conducted entirely in a BSL-2 facility.
   2. All rDNA technology studies involving BSL-2 organisms and/or BSL-2 host vector systems must be conducted in an RRI and approved by the IBC prior to experimentation, where applicable.
   3. All genome editing studies that include alteration of germline cells, insertion of gene drives, use of rapid trait development systems (RTDS®), etc., should be categorized as a BSL-2 study and must be conducted at an RRI and approved by the IBC from the institution. Qualified scientists are expected to ensure that student research protocols address appropriate intrinsic and extrinsic containment containment precautions.

Documentation and Approval:

1. The student and all of the adults involved in a research project must conduct and document a risk assessment on Form (6A) to define the potential level of harm, injury or disease to plants, animals and humans that may occur when working with biological agents. The risk assessment determines a biosafety level which in turn determines if the project can proceed, and if so, the proposed laboratory facility is properly equipped and all personnel are trained and appropriate supervision is planned:
   1. Give source of the organism and describe BSL assessment process and BSL determination.
   2. Detail safety precautions and discuss methods of disposal.
2. Prior review and approval is required for the use of potentially hazardous microorganisms (including bacteria, viruses, viroids, rickettsia, fungi, and parasites) and recombinant DNA (rDNA) technologies.
   1. 1. An affiliated fair SRC, an IBC or an IACUC must approve all research before experimentation
      2. The initial risk assessment determined by the student researcher and adults supervising the project must be confirmed by the SRC, IBC or IACUC.
3. Any proposed changes in the Research Plan/Project Summary by the student after initial local or affiliated fair SRC approval must undergo subsequent SRC, IBC or IACUC review and approval before such changes are made and before experimentation resumes.
4. The following forms are required:
   1. Checklist for Adult Sponsor (1)
   2. Student Checklist (1A)
   3. Research Plan/Project Summary
   4. Approval Form (1B)
   5. Regulated Research Institution Form (1C) — when applicable
   6. Qualified Scientist (2), when applicable
   7. Risk Assessment (3), when applicable
   8. PHBA Risk Assessment Form (6A), when applicable
   9. The BSL-2 Checklist when a BSL-2 facility is used that is not at a Regulated Research Institution.

Exempt Studies (No SRC Pre-Approval Required):

The following types of studies are exempt from prior SRC review, but require a Risk Assessment Form 3:

• Studies involving protists and archaea
• Research using manure for composting, fuel production, or other non-culturing experiment
• Commercially available color change coliform detection test kits; these kits must remain sealed and must be properly disposed
• Studies involving decomposition of vertebrate organisms (such as in forensic projects)
• Studies with microbial fuel cells in which the device is sealed during experimentation and disposed of properly at the conclusion of the study
• Studies involving fermentation of baker’s yeast and brewer’s yeast, except in rDNA studies
• Studies involving Lactobacillus, Bacillus thuringiensis, nitrogen-fixing, oil-eating, and algae-eating bacteria introduced into their natural environment (not exempt if cultured in a petri dish environment)
• Studies involving water or soil microbes not concentrated in media conducive to their microbial growth
• Studies of mold growth on food items if the experiment is terminated at the first evidence of mold
• Studies of slime molds and edible mushrooms
• Studies involving coli OP-50 and other strains of E. coli that are used solely as a food source for C. elegans and are performed at school and are not subject to additional rules for recombinant DNA studies or use of antibiotic-resistant organisms
• Studies involving coli K-12 that are performed at school and are not subject to additional rules for recombinant DNA studies or use of antibiotic resistant organisms