The official Biosafety in Microbiological and Biomedical Laboratories guidelines[edit | edit source]
The "bible" on biosafety guidelines in the US is the CDC's "Biosafety in Microbiological and Biomedical Laboratories" (BMBL), currently in its 5th Edition. The section on Biosafety Level 1 (BSL-1) starts on page 30, and is copied below:
Biosafety Level 1 is suitable for work involving well-characterized agents not known to consistently cause disease in immunocompetent adult humans, and present minimal potential hazard to laboratory personnel and the environment. BSL-1 laboratories are not necessarily separated from the general traffic patterns in the building. Work is typically conducted on open bench tops using standard microbiological practices. Special containment equipment or facility design is not required, but may be used as determined by appropriate risk assessment. Laboratory personnel must have specific training in the procedures conducted in the laboratory and must be supervised by a scientist with training in microbiology or a related science.
The following standard practices, safety equipment, and facility requirements apply to BSL-1.
Standard Microbiological Practices[edit | edit source]
- The laboratory supervisor must enforce the institutional policies that control access to the laboratory.
- Persons must wash their hands after working with potentially hazardous materials and before leaving the laboratory.
- Eating, drinking, smoking, handling contact lenses, applying cosmetics, and storing food for human consumption must not be permitted in laboratory areas. Food must be stored outside the laboratory area in cabinets or refrigerators designated and used for this purpose.
- Mouth pipetting is prohibited; mechanical pipetting devices must be used.
- Policies for the safe handling of sharps, such as needles, scalpels, pipettes, and broken glassware must be developed and implemented. Whenever practical, laboratory supervisors should adopt improved engineering and work practice controls that reduce risk of sharps injuries. Precautions, including those listed below, must always be taken with sharp items. These include:
- Careful management of needles and other sharps are of primary importance. Needles must not be bent, sheared, broken, recapped, removed from disposable syringes, or otherwise manipulated by hand before disposal.
- Used disposable needles and syringes must be carefully placed in conveniently located puncture-resistant containers used for sharps disposal.
- Non-disposable sharps must be placed in a hard walled container for transport to a processing area for decontamination, preferably by autoclaving.
- Broken glassware must not be handled directly. Instead, it must be removed using a brush and dustpan, tongs, or forceps. Plastic ware should be substituted for glassware whenever possible.
- Perform all procedures to minimize the creation of splashes and/or aerosols.
- Decontaminate work surfaces after completion of work and after any spill or splash of potentially infectious material with appropriate disinfectant.
- Decontaminate all cultures, stocks, and other potentially infectious materials before disposal using an effective method. Depending on where the decontamination will be performed, the following methods should be used prior to transport:
- Materials to be decontaminated outside of the immediate laboratory must be placed in a durable, leak proof container and secured for transport.
- Materials to be removed from the facility for decontamination must be packed in accordance with applicable local, state, and federal regulations.
- A sign incorporating the universal biohazard symbol must be posted at the entrance to the laboratory when infectious agents are present. Posted information must include: the laboratory’s biosafety level, the supervisor’s name (or other responsible personnel), telephone number, and required procedures for entering and exiting the laboratory. Agent information should be posted in accordance with the institutional policy.
- An effective integrated pest management program is required. (See Appendix G.)
- The laboratory supervisor must ensure that laboratory personnel receive appropriate training regarding their duties, the necessary precautions to prevent exposures, and exposure evaluation procedures. Personnel must receive annual updates or additional training when procedural or policy changes occur. Personal health status may impact an individual’s susceptibility to infection, ability to receive immunizations or prophylactic interventions. Therefore, all laboratory personnel and particularly women of childbearing age should be provided with information regarding immune competence and conditions that may predispose them to infection. Individuals having these conditions should be encouraged to self-identify to the institution’s healthcare provider for appropriate counseling and guidance.
Special Practices[edit | edit source]
Safety Equipment (Primary Barriers and Personal Protective Equipment)[edit | edit source]
- Special containment devices or equipment, such as BSCs, are not generally required.
- Protective laboratory coats, gowns, or uniforms are recommended to prevent contamination of personal clothing.
- Wear protective eyewear when conducting procedures that have the potential to create splashes of microorganisms or other hazardous materials. Persons who wear contact lenses in laboratories should also wear eye protection.
- Gloves must be worn to protect hands from exposure to hazardous materials. Glove selection should be based on an appropriate risk assessment. Alternatives to latex gloves should be available. Wash hands prior to leaving the laboratory. In addition, BSL-1 workers should:
- Change gloves when contaminated, glove integrity is compromised, or when otherwise necessary.
- Remove gloves and wash hands when work with hazardous materials has been completed and before leaving the laboratory.
- Do not wash or reuse disposable gloves. Dispose of used gloves with other contaminated laboratory waste. Hand washing protocols must be rigorously followed.
Laboratory Facilities (Secondary Barriers)[edit | edit source]
- Laboratories should have doors for access control.
- Laboratories must have a sink for hand washing.
- The laboratory should be designed so that it can be easily cleaned. Carpets and rugs in laboratories are not appropriate.
- Laboratory furniture must be capable of supporting anticipated loads and uses. Spaces between benches, cabinets, and equipment should be accessible for cleaning.
- Bench tops must be impervious to water and resistant to heat, organic solvents, acids, alkalis, and other chemicals.
- Chairs used in laboratory work must be covered with a non-porous material that can be easily cleaned and decontaminated with appropriate disinfectant.
- Laboratories windows that open to the exterior should be fitted with screens.
Biosafety level 1 (BSL1) guidelines for teaching laboratories[edit | edit source]
Based on the American Society for Microbiology's Guidelines for Biosafety in Teaching Laboratories.
Preamble: Educators need to be aware of the risks inherent in using microorganisms in the laboratory and must use best practices to minimize the risk to students and the community. The following guidelines are designed to encourage awareness of the risks, promote uniformity in best teaching practices, and protect the health and wellness of our students. These guidelines are not mandatory, but are designed to promote best practices in the teaching laboratory. Note that not all institutions are equipped to handle organisms in a BSL2 setting. Work with microbes at the K-12 level, informal education settings (e.g., science fairs, museums, science centers, camps, etc.), and in undergraduate non-microbiology laboratories would almost always be at BSL1. Even though organisms manipulated in a BSL1 laboratory pose a low level of risk to the community and are unlikely to cause disease in healthy adults, most of the microorganisms used in the microbiology teaching laboratory are capable of causing an infection given the appropriate circumstances. Many best practices should be adopted to minimize the risk of laboratory-acquired infections and to train students in the proper handling of microorganisms. The practices set forth in these guidelines fall into six major categories: personal protection, laboratory physical space, stock cultures, standard laboratory practices, training, and documents. For ease of use, the requirements and practices are brief. Explanatory notes, sample documents, and additional resources are in the appendix.
Personal Protection Requirements[edit | edit source]
- Wear safety goggles or safety glasses when handling liquid cultures, when performing procedures that may create a splash hazard, or when spread plating.
- Wear closed-toe shoes that cover the top of the foot.
- Wear gloves when the student’s hands have fresh cuts or abrasions, when staining microbes, and when handling hazardous chemicals. Gloves are not required for standard laboratory procedures if proper hand hygiene is performed. Proper hand hygiene involves thorough hand cleansing prior to and immediately after finishing handling microorganisms and any time that microbes accidentally contact the skin. Hand cleansing is performed by washing with soap and water or rubbing with an alcohol-based hand sanitizer.
- Recommended: Wear laboratory coats.
Laboratory Physical Space Requirements[edit | edit source]
- Require all laboratory space to include:
- Nonporous floor, bench tops, chairs, and stools.
- Sink for hand washing.
- Eyewash station.
- Lockable door to the room.
- Follow proper pest control practices.
- Recommended: Keep personal belongings in an area separate from the work area.
- Recommended: Use a working and validated autoclave.
Stock Culture Requirements[edit | edit source]
- Only use cultures from authorized, commercial, or reputable sources (e.g., an academic laboratory or state health department). Do not subculture unknown microbes isolated from the environment because they may be organisms that require BSL2 practices and facilities.
- Maintain documents about stock organisms, sources, and handling of stock cultures.
- Obtain fresh stock cultures of microorganisms annually (e.g., purchased, revived from frozen stock cultures, etc.) to be certain of the source culture, minimize spontaneous mutations, and reduce contamination.
Standard Laboratory Practices[edit | edit source]
- Wash hands after entering and before exiting the laboratory.
- Tie back long hair.
- Do not wear dangling jewelry.
- Disinfect bench before and after the laboratory session with a disinfectant known to kill the organisms handled.
- Use disinfectants according to manufacturer instructions.
- Do not bring food, gum, drinks (including water), or water bottles into the laboratory.
- Do not touch the face, apply cosmetics, adjust contact lenses, or bite nails
- Do not handle personal items (cosmetics, cell phones, calculators, pens, pencils, etc.) while in the laboratory.
- Do not mouth pipette.
- Label all containers clearly.
- Keep door closed while the laboratory is in session. Laboratory director or instructor approves all personnel entering the laboratory.
- Minimize the use of sharps. Use needles and scalpels according to appropriate guidelines and precautions.
- Use proper transport vessels (test tube racks) for moving cultures in the laboratory, and store vessels containing cultures in a leak-proof container when work with them is complete.
- Use leak-proof containers for storage and transport of infectious materials.
- Arrange for proper (safe) decontamination and disposal of contaminated material (e.g., in a properly maintained and validated autoclave) or arrange for licensed waste removal in accordance with local, state, and federal guidelines.
- Do not handle broken glass with fingers; use a dustpan and broom.
- Notify instructor of all spills or injuries.
- Document all injuries according to school, university, or college policy.
- Use only institution-provided marking pens and writing instruments.
- Teach, practice, and enforce the proper wearing and use of gloves.
- Advise immune-compromised students (including those who are pregnant or may become pregnant) and students living with or caring for an immune-compromised individual to consult physicians to determine the appropriate level of participation in the laboratory.
- Recommended: Keep note-taking and discussion practices separate from work with hazardous or infectious material.
- Recommended: Use microincinerators or disposable loops rather than Bunsen burners.
Training Practices[edit | edit source]
- Be aware that student assistants may be employees of the institution and subject to OSHA, state, and/or institutional regulations.
- Conduct extensive initial training for instructors and student assistants to cover the safety hazards of each laboratory. The institution’s biosafety officer or microbiologist in charge of the laboratories should conduct the training.
- Conduct training for instructors whenever a new procedural change is required.
- Conduct training for student assistants annually.
- Require students and instructors to handle microorganisms safely and responsibly.
- Inform students of safety precautions relevant to each exercise before beginning the exercise.
- Emphasize to students the importance of reporting accidental spills and exposures.
Document Practices[edit | edit source]
- Require students to sign safety agreements explaining that they have been informed about safety precautions and the hazardous nature of the organisms they will handle throughout the course.
- Maintain student-signed safety agreements at the institution.
- Prepare, maintain, and post proper signage.
- Document all injuries and spills; follow school/college/university policy, if available.
- Make Material Safety Data Sheets (MSDS) available at all times; follow institutional documentation guidelines regarding number of copies, availability via print or electronic form, etc.
- Post emergency procedures and updated contact information in the laboratory.
- Maintain and make available (e.g., in a syllabus, in a laboratory manual, or online) to all students a list of all cultures (and their sources) used in the course.