Experiment 2- Lab Safety Part A

*The following information is from your lab manual. You will be able to complete all of WEEK 1's assignments without having the manual OR kit. For Week 2's assignments, you will HAVE TO HAVE THE KIT AND MANUAL. I will no longer put information from either on Canvas after Week 1.

Introduction

Welcome to the Introductory Microbiology Lab Manual!

Microbiology is the study of microorganisms. Microorganisms, or microbes, include any organism that is too small to be seen by the unaided eye. Most of the microorganisms that we encounter are not harmful to us. In fact, many are quite beneficial to us and our surrounding community. Microorganisms are used in industrial production applications as well as employed to solve difficult environmental problems, such as oil spills.

Of course, there are a number of microorganisms that have harmful effects to humans, animals, plants, and even other microorganisms! Some microbes cause people or animals to get sick; or, even have a destructive effect on the environment. It is for this reason that there are defined safety procedures to follow and equipment to use when working with microorganisms.

Playing a game is more enjoyable than watching or reading about it. The same can be said for learning about microbiology. Performing experiments using microorganisms that are common in our environment is fun and informative. Throughout the experiments in this lab, you will come into contact with a variety of different microbes. While you will not be supplied with infectious organisms, you will be asked to isolate and culture (grow) microbes from your surrounding environment. It is therefore critically important to learn how to properly work with microbes to ensure your safety.

This manual is written for performing microbiology experiments using common microbes that can be readily isolated from every day materials, such as soil, food, the bottoms of shoes, from inside the mouth, etc. Microbes from these sources are generally much less hazardous than those that are used in a traditional microbiology lab setting. The experiments in this lab manual do not use dangerous human pathogens (disease causing organisms). However, many microbes are potentially dangerous, meaning that they would not normally cause disease in a healthy individual but that they could cause disease if they were introduced (for example by an open wound, cut, or open mucus membrane)  an immunocompromised individual. Generally speaking, immunocompromised individuals are people who have immune systems working at decreased or abnormal levels. This makes the person more susceptible o infections, disease, etc. If you have any concerns about your ability to perform these experiments safely, please consult your health care provider or instructor.

In addition to working with microorganisms, you will be working with chemicals, glassware, open flames, and sharp objects that can be hazardous if used carelessly or improperly. You should always use the safety goggles, gloves, and other personal protective equipment provided in your kit.

 

Safety Procedures, Practices and Rules

  • • Wash your hands with soap before beginning and after completing each lab.
  • • Do not eat, drink, chew gum, or smoke while performing the experiments.
  • • Do not put anything (pens, pencils, fingers, etc.) in your mouth while performing these experiments.
  • • If you are performing these experiments in your home, prepare a work space that is free of any
  • food or drinks.
  • • Disinfect your work area with isopropyl alcohol or 10% bleach solution both before and after
  • performing the experiments.
  • • Extinguish flame(s) before using or spreading isopropyl alcohol.
  • • Wear the safety equipment (see below) that is provided in the kit.
  • • Always read the experiment thoroughly before beginning it.
  • • Tie back long hair.
  • • Use a broom and dustpan to collect broken glass, place it in newspaper, and dispose of properly.
  • Never pick up broken glass with bare hands.
  • • When using a flame source, ensure that the area is free of loose paper, gas, and any other
  • flammable or combustible material. Ensure that the flame is extinguished when you are finished
  • with it.obiology Lab Safety• Material Safety Data Sheets (MSDS) for all chemicals provided are located on our website
  • (www.esciencelabs.com). These sheets contain all the necessary information regarding the danger, safe use, and disposal of each chemical.

 

Safety Equipment

Safety Goggles/Glasses-

Safety goggles and glasses are used to protect your eyes and should be worn at all times while

doing these experiments, even if you are not actively working with microor ganisms.

Gloves-

Gloves serve two purposes when working with microorganisms. Gloves protect your hands from

exposure to microorganisms and limit the spread of microorganisms present on your skin to the

cultures with which you are working. It is very important not to touch your work area with gloves

that have been contaminated with microbes. You should change your gloves before performing

other procedures. Gloves should always be worn while doing these experiments.

Lab Coat/Apron-

A lab coat or apron limits your exposure to the microbes you are working with. It also limits exposing the cultures you are working with to the microorganisms present on your skin. You should wear a lab coat every time you perform an experiment. Alternatively, a clean, long sleeved shirt that buttons or

snaps shut can be worn in place of a lab coat. The shirt must fully cover your arms and be removable without pulling it over your head. Collectively, safety goggles or glasses, gloves, and a lab coat or apron comprise the personal protective equipment that will be used in completing the experiments inthis manual.

Safety Shower-

Safety showers are used if someone/thing is exposed to a large quantity of microorganism(s). They

are also used if a hazardous chemical is spilled on a person and that person is unable to rinse it off

thoroughly in a sink. They may even be used in the event that clothes or an object catches fire. A

standard home shower sufficiently serves as a safety shower should the need arise.

Eye Wash-

An eye wash is used if your eyes are exposed to a microorganism or if a harmful chemical is

splashed into your eyes or face. In the home, use the nearest sink and flush your eyes or face with

cool water for at least 20 minutes. If the sink has a sprayer, use it to rinse your eyes or face while

making sure the water drains into the sink.

Fire Extinguisher-

A fire extinguisher can be used to put out small to medium sized fires. Direct the fire extinguishing

material at the base of the fire to properly put it out. A home fire extinguisher should be rated for A, B,

and C type fires. This is clearly marked on the outside of the extinguisher.

Laboratory Fume Hood-

A laboratory fume hood removes harmful gases and fumes that may be present when doing an experiment. You should always use a fume hood when working with corrosive, noxious, or flammable materials. Chemicals used in this kit will not require the use of a fume hood.

Biological Safety Cabinet-

A biological safety cabinet (BSC) is used as the main barrier to prevent exposure to infectious microorganisms in a laboratory setting. A BSC is not the same as a fume hood. Airflow in a BSC is partially recirculated to the room in which a BSC is housed. Additionally, a BSC contains a high efficiency particulate air (HEPA) filter which traps particulate (i.e., microorganisms), but not nonparticulate (i.e., fumes) matter. A BSC will not be required to complete the experiments in this manual.

 

Biological Safety Levels

The Centers for Disease Control and Prevention (CDC), in cooperation with the National Institutes of Health (NIH), have developed standards to offer protection when using biological pathogens. You are encouraged to review the publication (available here: http://www.cdc.gov/biosafety/publications/bmbl5/index.htm) which describes the practices, facilities, and safety equipment recommended for use in the four defined biosafety levels (BSL). A BSL describes the minimum required containment measures that should be used to ensure safety when working with various microorganisms. These measures are designed to reduce or eliminate exposure to potentially infectious microbes. The four BSL are outlined in Table 1.

 

BSL

Agents

Practices

Equipment

Facilities

1

Not   known to cause disease in healthy adults

Standard   microbiological

practices

None   required

Open   bench top Hand washing sink

2

Associated   with disease; main hazard is percutaneous injury, ingestion, and mucus

membrane   exposure

BSL-1   plus limited

access,   biohazard

warning   sign, sharps precautions, manual detailing decontamination

and   medical

surveillance   polices

Class   I or II BSC for procedures

that   generate

aerosols   or

splashes;   gloves, lab coat, goggles; respiratory

protection   if

needed

BSL-1   plus available

autoclave

3

Associated   with

potential   for aerosol

transmission;   disease may be lethal

BSL-2   plus controlled access; decontamination of all waste and lab clothing;   baseline serum drawn for each worker

Class   I or II BSC for all procedures;

gloves,   lab coat, goggles;

respiratory

protection   if

needed

BSL-2   plus physical

separation   from access points, selfclosing, double door access, no recirculated air,   negative airflow into BSL-3 lab

4

Associated   with

risk   of lifethreatening

disease,   aerosol

transmission   or unknown risk of

transmission

BSL-3   plus clothing

change   before entering lab, shower when leaving lab, all materials decontaminated

Class   III BSC or

Class   I or II if

used   with full

body,   independent

air   supply,

positive   pressure

suit

BSL-3   plus isolated labs, dedicated air supply and exhaust

**Only BSL-1 procedures will be used in these experiments!!!**

 

Examples of BSL-1 Organisms: Escherichia coli (most species), Bacillus subtilus, and other organisms commonly found in the local environment.These microorganisms are well characterized and not known to cause disease in healthy adults. No special laboratory equipment, including a BSC, is required. Work may be performed on an open bench and exposure is limited by the use of personal protective equipment: goggles, gloves, and a lab coat.

 

Examples of BSL-2 Organisms: Bacillus anthracis (anthrax), Bordetella pertussis (whooping cough), Clostridium botulinum (botulism), Human Immunodeficiency Virus (HIV).icrobiology Lab Safety

Examples of BSL-3 Organisms: Mycobacterium tuberculosis (tuberculosis), Salmonella typhi (typhoid fever), Yellow Fever Virus.

 

Examples of BSL-4 Organisms: Smallpox virus, Ebola virus, hemorrhagic fever virus.

Again, you will NOT be using any organisms that require BSL-2, BSL-3, or BSL-4 containment procedures.

 

 

If you have any doubt or concerns about safety:

• STOP!

• Double-check the manual

• Refer to our website: www.esciencelabs.com

• Email: Help@esciencelabs.com

• Call for help: 1-888-ESL-Kits (1-888-375-5487)