You are responsible for reading and understanding the content from the section Basic Microbiology Laboratory Safety.

Transcription

1 BIOSAFETY You are responsible for reading and understanding the content from the section Basic Microbiology Laboratory Safety. Additional Safety Information: The foundation of laboratory biosafety is the recognition of four Containment Levels. Specific practices and infrastructure requirements are established for each containment level. All work with microorganisms in a laboratory setting is assigned to a specific containment level based on the properties of the organism involved, and on the nature of the procedures to be used. The most comprehensive source of information on laboratory biosafety is Biosafety in Microbiological and Biomedical Laboratories (BMBL) 5th Edition 1. This document is the best starting point if you want to determine which containment level is appropriate for a given project and what equipment and procedures will be necessary to satisfy that level of containment. In this class, work is ordinarily conducted at Biosafety Level 1 (BSL-1), with agents that are categorized as Risk Group 1. These agents "not associated with disease in healthy adult humans" 1. In several experiments we will use bacteria that are classified as Risk Group 2, and are handled using BSL-2 protocols. These bacteria are defined as " agents associated with human disease that is rarely serious and for which preventive or therapeutic interventions are often available" according to the National Institutes of Health (NIH) Guidelines for Research involving Recombinant DNA Molecules. We will review the guidelinesabout working with BSL-2 organisms during the weekly discussion session prior to carrying out these specific experiments. There is additional information on the course web site. 1 Available for free download at the CDC website,

2 POLICIES OF METX119L The follow practices will be enforced rigorously in every lab: No eating, drinking, smoking, or applying cosmetics in the laboratory. Do not pipette by mouth. Notify the TA or the instructor if you have an open cut or burn. Notify the TA or instructor of any spilled cultures so that they can be properly cleaned up and decontaminated. Notify the TA or instructor about broken lab equipment and glassware. Broken glassware must not be handled directly. Instead, it must be removed using a brush and dustpan, tongs, or forceps. After each lab period, wipe your bench with disinfectant and wash your hands before you leave. Cultures, and materials contaminated with cultures of microorganisms, should be properly labeled and identified with the approved biohazard warning logo. It's important to dispose of all waste properly, as follows: Agar plates should be placed in a designated biohazard bag/container for autoclaving. Flasks and tubes containing cell suspensions should be placed in the autoclaving pan provided at the back of the class. Contaminated pipettes, swabs, kimwipes and micropipette tips are collected in appropriate biohazard containers on the benchtops for autoclaving. The prep staff will autoclave and discard these materials after each lab. Nothing that touched bacteria should go in the regular trash. You will get specific instructions for chemical disposal, or just ask. Follow instructions, protocols, and techniques as specified either in the handouts, discussed by the instructor, and demonstrated in class. Deviation from this may put you or others at risk for being exposed to biological and chemical agents. Perform all procedures carefully to minimize the creation of splashes and/or aerosols. There is a zero tolerance for improper conduct in the laboratory. Any student(s) who put themselves and others at risk will be immediately disciplined according to the Official University Policy on Academic Integrity for Undergraduate Students : nts/ CHEMICAL SAFETY Several of the laboratory exercises involve hazardous chemicals. The nature of the hazards will be identified and the necessary precautions described in the lab handouts and in pre-lab lectures.

3 Understand all cautionary notes in the written lab handouts before coming to class. Do not begin work until the TA or the instructor has discussed all issues and questions regarding safe handling and disposal of chemicals. PERSONAL PROTECTION Lab Coats: Protective laboratory coats, gowns, or uniforms are recommended to prevent contamination of personal clothing. Proper Clothing: You must wear closed-toed shoes, and clothing that substantially covers arms, legs, and feet are required. No shorts and flip flops. Eyes: 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 all the time. Safety goggles are provided in one of the lab drawers. Wear them whenever you are directed to do so in the lab handout or by the instructor. You can also bring your own safety glasses. Hands: Gloves must be worn to protect hands from exposure to hazardous materials. o Alternatives to latex gloves will be available. o Change gloves when contaminated, integrity has been compromised, or when otherwise necessary. o Remove gloves and wash hands when work with hazardous materials has been completed and before leaving the laboratory. Hand washing protocols must be rigorously followed. o Do not wash or reuse disposable gloves. Dispose of used gloves with other contaminated laboratory waste. o Do not leave the lab if you are wearing gloves. Eye Washes/Showers: Know the locations and uses of the eye wash fountain and safety shower. Fire: Find where the fire extinguisher, fire alarm, and nearest telephones are. Hazardous chemicals are never put down the drain or thrown in the ordinary trash. Appropriate waste containers will be available in the lab. You cannot enter the lab unless you are clothed properly. You can keep a pair of long pants and shoes in the lab--ask the Professor or TAs to help you find a location. ADDITIONAL HAZARDS UV: The germicidal lamp in the biological safety cabinet is a low pressure Hg arc that emits strongly in the short UV range of the spectrum. Exposure to short wave UV is mutagenic and will cause severe burns to exposed skin and damage your eyes if viewed directly. The glass window of the hood offers adequate protection for your eyes. However, the NIH recommends against the UV light being on whenever anyone is in the room (sash shut or not).

4 Flames: Bunsen burners are fire hazards. Turn them off whenever you are not using them. Know the location of the fire extinguisher and know when and how to use it. The extinguisher will be demonstrated in lab. Heavy things: The microscopes are heavy. Don't drop one on your foot. I HAVE READ AND UNDERSTOOD THE SAFETY GUIDELINES PRESENTED IN THIS DOCUMENT. I WILL ALSO READ AND UNDERSTAND OTHER SAFETY INFORMATION PRESENTED THROUGHOUT THE COURSE. PRINT YOUR NAME: SIGNATURE: DATE: (This statement should be saved in your lab notebook)

5 BIOSAFETY ASSIGNMENT This assignment is based on information in "Biosafety in Microbiological and Biomedical Laboratories (aka BMBL), 5th ed." (Go to the Online Resources section on the course web site and follow the link on the BMBL). Sections III, IV, VII and Appendix A have the information you will need, although the others are fascinating as well. Note: BSC is an abbreviation for Biological Safety Cabinet (other abbreviations in the last appendix). 1. Pick three pieces of Primary Personal Protective equipment, and describe what they are used for. 2. Describe two types of secondary barriers used in BSL-2 labs and what they are used for. 3. Describe three differences between how work is done in a BSL-2 lab as compared to a BSL-1 lab. 4. What BSL is Salmonella considered? How about Brucella? Make sure to look at all types. 5. What are the essential differences between a Type I, Type II, and Type III Biological Safety Cabinet? (See biosafety links on the course website) Is the cabinet near the lab (room 215A) a Type I, Type II, or Type III cabinet