How do I make my lab BSL-1 compliant?
I've not been able to find a definitive document or checklist that will help me establish a BSL-1 compliant lab, and I'd like to make sure that I am following correct procedures. If there's a straightforward 'how-to', I'd love to know where it is! I only want to do some simple PCR, bacterial culture from the environment, and microscopy, so I'm sure compliance isn't hard. Please point me in the right direction!
Answer from a Biosafety Officer:
March 8, 2014
A laboratory often handles a number of different types of hazards. The biological hazards are only one of the potentially dangerous things that can be found in a research laboratory. This often makes it quite difficult to figure out what things you need to be careful about because each hazard is regulated by different agencies.
The purpose for most of the regulations that have been implemented to date is to protect employees conducting this work and the environment. So if you think about everything that you do in these terms you will be on the right track.
Since this is DIYbio, we will address the biological hazards first. The intent here is to make sure that you don't create/amplify something in the lab that will be a hazard to you or others who might enter your lab. With that said there is a lot of bacteria in the soil and some of them are human pathogens.
For example, Bacillus anthracis and Pseudomonas aeruginosa, are commonly found in soils but are dangerous to people. As long as they remain in the soil they generally don't cause us problems because we don't come in contact with large numbers of them. However when you bring these into the laboratory and start culturing them the number of organisms increases and so does your risk. So you always need to be careful and deliberate about what you are trying to amplify from any sample.
So how do we deal with biological risk?
First we build our buildings to make sure that they have the right types of equipment to help us keep the things in the laboratory that are meant to stay in laboratory and dispose of other things properly. This is why you commonly see hand sinks at the door so that people can wash their hands prior to leaving the space. That way they don't carry contaminants into the environment. The criteria for building laboratory go hand-in-hand with practices and procedures meant to keep the laboratory safe. Of course as the level of risk increases so does the cost and the amount of equipment required to contain amplified organisms inside the lab.
Fortunately, in a BSL-1 laboratory (handling BSL-1 organisms) the risk is low and therefore the number of practices, procedures and building requirements are also not very stringent. However, if you culture and amplify soil samples you do not necessarily know that you are only going to be amplifying organisms that are generally handled safely in the BSL-1 environment. Back to my previous example of Pseudomonas aeruginosa and Bacillus anthracis, Pseudomonas aeruginosa is generally handled at BSL-2 because it is a known human pathogen. Bacillus anthracis is even more of a concern. In the US, possession, use, storage and\or transfer of this organism is regulated under the select agent rules. It is actually a federal crime to possess, use and store these organisms without being appropriately registered and certified by the CDC Select Agent Program.
There are good resources to look at when trying to assess risks associated with particular organisms. The CDC/NIH publication Biosafety in Microbiological and Biomedical Laboratories (BMBL) provides agent summaries for many bacterial, fungal, rickettsial and viral hazards as well as criteria for practices, equipment and laboratory design for each of the four biosafety levels. Another excellent resource is from the Public health agency of Canada. They have created pathogen safety data sheets the providing enormous amount of information regarding the hazards and handling requirements of these organisms.
Back to your question regarding BSL-1 laboratories, the BMBL describes standard microbiological practices, safety equipment, and laboratory facilities required to meet BSL-1 standards. In general, BSL-1 practices and procedures are 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.(1)
For specific requirements for BSL-1, refer to Section IV of the BMBL. Additionally, below is a list of useful standard practices for BSL-1 and above
- Your work area should be dedicated for work with microbes. Don’t store cultures or plates in your kitchen. Ideally, cultures should not be stored in living areas such as bedrooms.
- Always wash your hands after working with microbes and before leaving your work area.
- Do not eat, drink, apply cosmetics, or store food or toiletries in your work area.
- Don’t mouth pipet.
- Be cautious when using needles, scalpels, and other sharp items with infectious microbes. Limit their use, if possible. Avoid bending, shearing, breaking, or recapping needles. Make sure that you dispose of these items properly – sharps containers and return programs are available at some pharmacies. Be aware that state regulations may apply to disposal of contaminated needles or sharps.
- Use careful work practices to avoid splashes and production of aerosols.
- Decontaminate your work area when you are finished working. A simple mixture of 1 part bleach to 9 parts water is a very effective disinfectant. Make sure that all surfaces that you want to disinfect remain wet with the diluted bleach solution for at least 5-10 Minutes.
- Decontaminate your cultures and plates before disposal. Cultures should be treated with a 1:10 dilution of bleach (1 part bleach to 9 parts liquid culture e.g. 10 mL bleach in 90 mL liquid culture). The surfaces of petri dishes can be soaked in a 1:10 dilution of bleach for 2 hours before disposal (2).
- Proper Personal Protective Equipment includes wearing an outer layer of clothing that can be sanitized or disposed (disposable lab coat, disposable gown), latex or nitrile gloves, and eye protection if splashes of microbes or chemicals may occur. Don’t wash or reuse disposable gloves.
If you stick with these practices and procedures for your biological hazards you should be fine. Please remember that if you're working with chemicals you have to be careful what you sink dispose of these materials often end up in our watersheds. Again look to your material safety data sheets for information regarding the chemicals that you intend to use. If the chemicals did not come with one you can request one from the manufacturer so that you understand hazards to your self and the environment. Anything that is labeled as a: carcinogen, mutagen or teratogen in should not be disposed of in the sink and should not come in contact with you. Make sure that you wear gloves that are appropriate to the chemical that you are handling. Latex gloves do not provide adequate protection for many of the chemicals that you might handle in doing DNA extractions or amplification for PCR.
1. Wilson, D., and L. Chosewood, editors. Biosafety in Microbiological and Biomedical Laboratories (BMBL), 5th edition. Centers for Disease Control and Prevention and National Institutes of Health. 2009. http://www.cdc.gov/biosafety/publications/bmbl5/ (Accessed on 02/05/2014).
2. Public Health Agency of Canada. Pathogen Safety Data Sheets and Risk Assessment. http://www.phac-aspc.gc.ca/lab-bio/res/psds-ftss/index-eng.php (Accessed on 2/7/2014)