Developed for Science Laboratories

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General Laboratory Safety Training

• Developed for Science Laboratories
• September 2002

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• This training presentation has been created for
  those who are required to undergo general
  laboratory safety training and have not taken
  this training with their supervisor. It is
  also recommended for those laboratories that do
  not fall under regulation, but are needing a
  safety laboratory overview.

There are hyperlinks throughout the presentation.
Where there is the following icon additional
training is required.
Training
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Introduction

• Laboratory safety is the key to reducing injury
  and illness. There are many exposures in the
  laboratory that pose a hazard to your health and
  you may have never considered them as a hazard
  before. It is important to have proper training
  so you, as the employee, are aware of the
  potential dangers that may threaten your health
  or life.

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Introduction

• As you go through this training module, you will
  have a better understanding regarding the concept
  of safety and how safety is utilized in every
  aspect of your life. The University has an
  obligation to protect each student, assistant,
  faculty and staff. There are also regulations
  that pertain to ISU to ensure compliance. Lets
  take a look at the different agencies that are
  involved!

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Agency Information

• Several agencies are involved in safety
  compliance at the University. The following
  agencies impact our laboratories as far as safety
  is involved.

National Institute of Health (NIH)
In charge of laboratories that use or contain
recombinant DNA. They specify the practices for
constructing and handling the rDNA.
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Agency Information

• Centers for Disease Control and Prevention (CDC)

In charge of laboratories that use or contain
infectious agents. They are focused on
protecting personnel and the laboratory
environmental from exposure to infectious agents.
They are also aiming at preventative measures by
adhering to strict containment.
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Agency Information

• Occupational Safety and Health Administration
  (OSHA)

OSHA primarily monitors Hazardous Materials,
Hazard Communication, Bloodborne Pathogens, and
Occupational Exposure to Hazardous Chemicals in
Laboratories at ISU. OSHA is in charge of
monitoring work conditions and eliminating
physical and health hazards at the work place.
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Agency Information

• Environmental Protection Agency (EPA)

The EPA is in charge of the wastes that ISU
generates and stores. Many hazardous wastes are
used in research laboratories and they pose a
threat to humans, animals, plants, and the
environment. There are also biohazardous wastes
that are regulated and must be properly
decontaminated and disposed of.
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Agency Information

• Environmental Health Safety Office (EHS)

The EHS office plays a large role in overseeing
the various activities that take place on campus.
They interpret laws and regulations, and
maintain up-to-date records of current health and
safety standards. The duties range from fire
safety and laboratory safety to training and
emergency response.
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Why Training?

• Training is by far the most important aspect of
  safety. Training not only reduces illness/injury
  but also increases worker efficiency and
  awareness. Training is required by regulation
  and ISU has high regard for fulfilling this
  commitment. Preventing hazards and increasing
  awareness is the goal for the EHS office.

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Who is in Charge of Training?
Everyone on campus is in charge of training. The
individual is responsible for participation of
the training, comprehending the information, and
utilize the information for the specific duties.
The supervisor is in charge of providing proper
and effective training. The EHS office provides
assistance to departments in achieving regulatory
compliance and developing proactive strategies.

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Introduction to the Laboratory
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Lets Get Started!

• Working in a laboratory can be an exciting
  experience. It can also pose many threats and
  hazards that a traditional classroom does not.
  That is why it is important to know your
  surroundings. Know where the exits to your room
  are. There may be more than one exit which could
  be critical in the case of an emergency. Your
  supervisor will go over the emergency action plan
  including the escape route procedures for your
  room.

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Know Your Surroundings

• It is also recommended to be aware of the fire
  extinguishers in location to your laboratory. In
  order to fight a fire one must undergo the proper
  training. Campus personnel are highly encouraged
  to not fight fires. In the event of a fire, the
  first response is to evacuate the area and notify
  the fire department! Any campus phone will
  initially direct calls to the ISU Police Dept.
  and from there the fire dept. will be dispatched.
  From any campus phone just dial 911. Do not wait
  any longer than necessary to call, time is of
  essence!

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Know Your Surroundings

• Know where the fire alarm is in proximity to
  your laboratory. Is it right down the hall or is
  it in the stairwell? If there is a fire, a quick
  response is the best response. Have your
  supervisor show you the closest alarm. The fire
  safety training will detail the safety procedures
  for your lab.

Training
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Know Your Surroundings

• Many laboratories contain hazardous
  substances.
• A hazardous substance is defined as a
  material/substance that poses a physical or
  health hazard. This includes both chemicals and
  biological agents.
• A Biohazard is defined as any organism that is
  capable of replication and is capable of causing
  disease in human, animal or plant.
• There are differences between a physical hazard
  and a health hazard. Lets take a look.

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Know Your Surroundings

• A health hazard has the following
  characteristics

• Carcinogen
• Toxic or highly toxic
• Reproductive Toxins
• Irritants
• Corrosives
• Sensitizers
• Hepatotoxins
• Nephrotoxins
• Neurotoxins

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Know Your Surroundings

• A physical hazard has the following
  characteristics

• Explosive
• Flammable
• Oxidizer
• Pyrophoric
• Organic peroxide
• Compressed gas
• Combustible liquid
• Unstable (Reactive)
• Water-reactive

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Know Your Surroundings

• When physical hazards and health hazards exist,
  it is very important to know where the eye
  wash/safety shower is located. Unexpected
  accidents do occur and knowing where to go at the
  time of an emergency can reduce injury/illness.

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Know Your Surroundings

• First aid kits have a variety of quick relief
  items. If your lab has a first aid kit, find out
  where it is. If more than first aid is needed, it
  is recommended to go to Student Health Services
  for further treatment. In an event that would
  require more than first aid to be treated, report
  it to the EHS office within the next 24 hours.

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Know Your Surroundings

• When there are chemical, biological, or
  radioactive agents being used, an emergency spill
  kit should be available. If there is a spill kit
  in your lab, find its location. Further Spill
  information will be addressed later in this
  presentation.

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Know Your Surroundings

• Each laboratory has a telephone in a designated
  area for use. The emergency contact numbers are
  posted near the phone in every laboratory on ISU
  campus.

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Know What Hazards are Present
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Hazards in your Lab

• Each lab is faced with different hazards. There
  could be exposure to biological, chemical, or
  radioactive material, which may pose a variety of
  physical and/or health hazards. This training
  will be primarily focusing on biological hazards
  and some chemical hazards. Lets look at these
  hazards a little closer.

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Hazards in your Lab
A biological hazard includes an organism or
material of biological origin that could
potentially cause harm to humans, animals, or
plants.

• An infectious agent is an organism capable of
  producing infection or disease in human, animal
  or plant.
• Recombinant DNA (rDNA) is defined as molecules
  that are constructed outside living cells by
  joining natural or synthetic DNA segments to DNA
  molecules that can replicate in a living cell or
  molecules that result from the replication of
  those described above.

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Hazards in your Lab

• Infectious agents pose a threat because these
  agents can cause illness or death to both people
  and animals. Special precautions must be taken
  to reduce the potential release of these agents.
  Each laboratory that is using an infectious agent
  must perform an additional lab specific training.
  This is to communicate the hazards of the agent
  that is specific to your laboratory.

Training
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Hazards in your Lab

• Infectious agents fall under CDC guidelines.
  Depending on the organism you are working with or
  exposed to will depend on the proper safety
  precautions to take. These are designated at
  different Biosafety Levels (BSL). The more
  pathogenic the agent, the more stringent the
  safety precautions are.

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Hazards in your Lab

• The lab that works with infectious agents,
  depending on the Biosafety Level (BSL), will vary
  in accordance to the safety equipment that is
  used, the facility (lab) design, the equipment
  that is used, and the practices that must be
  followed.

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Hazards in your Lab

• Biosafety levels will be discussed in detail by
  your supervisor during the Lab Specific Training.

Training
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Hazards in your Lab

• rDNA can also poses a threat to human and animal
  health. Depending on the molecules that are
  being manipulated, the hazards will vary. rDNA
  can be manipulated using cultures, animals,
  plants, and also humans.
• The risk will depend on several factors
  including the organism and the media being used.
  The containment levels will depend on the
  organisms that carry the recombinant DNA.

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Hazards in your Lab

• The term containment in the laboratory setting
  is defined as the confinement of a biohazardous
  agent that is being cultured, stored,
  manipulated, transported, or destroyed in order
  to prevent or limit its contact with people
  and/or the environment. Methods used for
  containment include physical and biological
  barriers and inactivation using physical or
  chemical means.

Containment is integrated into the distinguished
biosafety levels.
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Hazards in your Lab

• Chemicals can pose a significant hazard. They
  should be limited to the use under a properly
  working fume hood. Chemicals can release
  hazardous fumes which not only harm the
  environment, but they can be a major health
  threat. They must be handled carefully and
  disposed of properly.

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Hazards in your Lab

• When a chemical is in the laboratory, the
  hazards of that chemical must be communicated to
  you. According to Occupational Safety and Health
  Administration (OSHA), a Chemical Hygiene Plan
  (CHP) is required to relay information regarding
  procedures, equipment, PPE, and work practices
  that are capable of protecting employees from
  health hazards.
• Your supervisor is in charge of providing the
  information contained in the CHP to you.

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Lab Safety

• The following guidelines have been established
  to minimize the hazards in a laboratory setting.
  It is important to take responsibility for your
  actions and to keep in mind that irresponsible
  acts could have lasting future effects.
• The next portion of the training is simply
  review and is meant to remind you of responsible
  conduct in a laboratory setting. As the hazards
  increase, the risks increase, and the
  responsibility must increase.

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Lab Attire
You should remember the following

• No open-toed shoes
• No shorts unless a lab coat is used
• Restrain hair when working with hazardous
  materials
• Remove protective clothing in public
• Use the proper Personal Protective Equipment for
  the job

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Personal Habits
Personal habits play a large role in minimizing
hazards. The following measures must be taken

• Do not eat, drink, smoke, chew gum or apply
  cosmetics, or remove/insert contact lenses while
  in the laboratory
• Do not store food or beverages in the lab or in
  chemical refrigerator
• Do not mouth pipette
• Wash hands before leaving laboratory or after
  handling contaminated material

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Safe Practices

• These safe practices should be followed to
  ensure safe working conditions
• Do not use chipped or cracked glassware
• When working with hazardous materials, have a
  second person nearby
• Know emergency procedures
• Keep the laboratory neat and clean
• Use hazardous chemicals under a fume hood and
  biohazardous materials under a biosafety cabinet
  (BSC)
• Decontaminate as needed
• All procedures should be performed to minimize
  aerosol

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• Know details/location of your vital Laboratory
  Information!

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Laboratory Information

• Each lab contains an Emergency Action Plan
  which is required for emergency situations. This
  is used to inform faculty/staff/students of the
  procedures to follow in the event of an
  emergency. This includes the following
  information

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Laboratory Information

• The labs that are using infectious agents
  and/or rDNA are also required to have the ISU
  Biosafety Manual available. Specific guidelines
  for research are outlined in this manual. If any
  questions arise regarding regulations, this
  manual should be used as a resource.

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Laboratory Information

• The Chemical Hygiene Plan (CHP) is similar to
  the Hazard Communication Program, but this is
  specifically for laboratories with chemicals.
  This is to relay information regarding
  procedures, equipment, PPE, and work practices
  that are capable of protecting employees from
  health hazards. If you are working with acids, it
  would be nice to know what personal protective
  equipment is necessary and what health hazards
  are associated with it.

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Chemical Hygiene
According to the laboratory standard, for those
labs that have chemicals the following training
topics must be addressed

• Methods and observations that may be used to
  detect the presence or release of a hazardous
  chemicals.
• Physical and health hazards
• Measures employees can take to protect themselves
  from hazards
• Details of the CHP

This training is covered in the Hazard
Communication Training Presentation
Training
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Chemical Hygiene Plan

• Methods and observations used to detect the
  presence or release of chemicals will be specific
  to your lab. A good indication of the presence
  of a chemical is to rely on your senses. Can you
  see it or smell it? Is a monitoring device
  needed to detect it? This can be important
  information if the chemical is an asphyxiant and
  pushes oxygen out of the room you are in.

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Chemical Hygiene

• Protection against chemicals is a combination of
  work procedures or practices, emergency
  procedures, and PPE. Some chemicals can only be
  handled under certain conditions. It is
  important to use proper handling procedures and
  practices as advised. The emergency procedures
  for chemical accidents is to first evacuate the
  area and then notify your supervisor, ISU campus
  police and EHS office if necessary.

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Chemical Hygiene

• Each lab that has chemicals will have a lab
  specific Chemical Hygiene Plan. This plan will
  detail the procedures and practices for your
  specific location. It is your responsibility to
  make sure that you review the CHP.

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Labeling

• It is important to know as much about a chemical
  as possible. The most dangerous substance is the
  one that has no label. Communicating information
  is essential in the science field.

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Labeling
Labeling is also required for chemicals that are
in a secondary container unless it is to be used
within an 8 hr period. When labeling the
secondary container the following information
must be included

• Identity of the hazardous chemical
• Appropriate hazard warnings
• Name and address of the chemical
  manufacturer, importer, or other responsible
  party.

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MSDS
There are several constituents that are covered
in the Laboratory Standard. Among these
constituents includes labeling and the MSDS.
Lets look more closely at the content of the
MSDS.
MSDS
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MSDS

• In addition to labeling in a laboratory, the
  next most important type of communication
  regarding hazards is the MSDS. This is the
  acronym for Material Safety Data Sheet. This
  will communicate the information necessary
  regarding hazards associated with chemicals and
  also biological agents.

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MSDS

• So what is an MSDS? An MSDS is a document that
  relays vital information about certain chemicals
  and biological agents. Check below to see what
  content the MSDS contains!

MSDS Content Click here!
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Reading the MSDS

• Each component of the MSDS is broken down into
  sections. Each section goes into detail about
  what precautions to take and the characteristics
  of the substance. This is for both chemicals and
  biological agents. See the below links for
  example MSDSs.

MSDS Chemical
MSDS Biological Agent
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Laboratory Information

• Why is an MSDS important? When you know
  characteristics about a substance it can aid in
  precautionary measure to take when using it.
  Also, if there is a spill either on a surface or
  on your skin, the MSDS can supply you with the
  information needed for first aid.

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MSDS

• The MSDS to every chemical in your lab must be
  available to you. It may be in a notebook in
  your lab. Make sure you find the location of the
  MSDSs in your room.

MSDS
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• Hazard Communication

If chemicals are used, Hazard Communication
training must be conducted. This training is
specifically for the labs that use chemicals and
the training can be accessed through the EHS
website.
Training
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Exposure Control Plan

• The Exposure Control Plan is required for the
  labs that have exposure to human blood or Other
  Potentially Infectious Material (OPIM). This
  includes human blood or human blood products such
  as semen or vaginal secretions. This also
  includes fluids such as cerebrospinal, synovial,
  pleural, pericardial, peritoneal and amniotic.
  Saliva and body fluid that is visibly
  contaminated with blood also is considered OPIM.
  

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Exposure Control Plan
The Exposure Control Plan is established to make
an exposure determination and a schedule of
implementation for methods of compliance, HBV
vaccination and post-exposure evaluation
including follow-up, HazCom, and recordkeeping.
If you are working with human blood or OPIM, you
must talk with your supervisor and locate the
Exposure Control Plan. There would only be a
select few laboratories that this standard would
apply to.

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Personal Protective Equipment
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What is PPE?
PPE is short for personal protective equipment.
This is the equipment that is necessary to
protect yourself from hazardous and biohazardous
materials. PPE could be gloves, safety glasses,
lab coat, shoe covers, respirator or any other
item that could protect you from dangerous
materials that you may encounter in the lab.

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When to use PPE
Knowing what to use and when to use it is the key
to properly protecting yourself. There could be
situations that would be more of a risk and
require more PPE than others. The next few
slides will help you in determining what you
should be using!

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When to use PPE

• Chemical usage poses a variety of hazards. They
  can be flammable, corrosive, even toxic just to
  name a few. Taking all precautions to avoid
  physical and/or health problems is the number
  one goal. You can never be too cautious!

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Chemicals PPE
When chemicals are being used there is always the
possibility of splashing. The proper PPE to use
when chemicals are involved would include

• Safety glasses
• Gloves
• Respirator (depending on the chemical and the
  exposure duration)
• The Permissible Exposure Limit (PEL) for an 8
  hr. shift can be obtained from the PEL/BEL
  reference guide. Contact EHS if questions
  arise.

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When to use PPE

• Biological agents as well as rDNA can pose
  serious health problems. Pathogens can cause
  disease and possible death in animals and humans.
  Containment of the agent/material and proper
  handling techniques are the best methods for
  minimizing risks that are associated with
  biological hazards. rDNA can also be a threat
  depending on the host and the vector that is
  being used.
• PPE is only one way that you can protect
  yourself. It is your responsibility to ensure
  that the proper PPE is used.

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Biological Agents rDNA PPE

• When biological agents and/or rDNA are being
  used the following PPE must be used

• Safety glasses
• Gloves
• Lab coat
• Respirator (Depending on the agent and whether or
  not an aerosol is created)

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When to use PPE

• Animals have the ability to bite and scratch
  unlike many of the other hazards. This can be of
  concern when the animal is infected and can
  transmit disease to a human. This is called
  zoonotic. To avoid infection, proper handling
  procedures must be utilized. The appropriate
  handling in combination with the PPE can help to
  reduce risk.

www.animalpicturesarchive.com
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Animals PPE

• When animals are being used the following PPE
  must be used

• Gloves-May need a variety to prevent animal
  bites.
• Lab coat-Needed to protect clothing and used as a
  barrier. To prevent the pathogen from leaving the
  lab, the lab coat must be worn and must not leave
  the lab.
• Safety glasses-Must be worn to prevent possible
  injury to the eyes by scratching or flying saw
  dust bedding. Splashing or aerosol also requires
  glasses.
• Respirator-May be needed depending on the
  pathogen. Check with safety office.
• Shoe covers-Used in the animal room and labs that
  work with biological agents.

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When to use PPE

• Arthropods are associated with potential risks
  when they escape since some are vectors of
  infectious human diseases. Precautions need to
  be taken to minimize the hazards associated with
  arthropods. Communication during lab specific
  training can help you understand the potential
  risks associated with the arthropod that you come
  in contact with.

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Arthropods PPE

• Depending on the arthropod that you encounter in
  the lab, the PPE could vary. In most cases the
  PPE below should be used used.

• Gloves
• Safety glasses (Use if arthropod is infected or
  if there is a chance of the arthropod
  spraying or jumping in your eye)
• Lab coat (If arthropod is known to be infected)

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• Proper Use of Personal Protective Equipment (PPE)

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Use of PPE

• Knowing how to properly use PPE can be the key
  to adequate protection. Not only do you want to
  make sure it is the proper size for you, but also
  make sure you are wearing it properly. If it is
  too big or too small, it is not right for you!
  Let your supervisor know if you need a different
  size.

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Use of PPE

• The main piece of equipment that must be used
  appropriately is the respirator. If you work
  with infectious agents, a respirator may be
  required depending on the biosafety level (BSL)
  of the agent. The more pathogenic the agent, the
  more PPE that is required. If a respirator is
  required, you will be part of the ISU Respiratory
  Protection Program. This requires a physical,
  fit-test, and additional training before use. If
  the respirator is not properly fit to you,
  serious health problems can result.

Training
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Use of PPE

• If you think you need a respirator talk with
  your supervisor or contact EHS (438-8325)

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Location Availability of PPE

• Each Lab should have a designated area for the
  PPE that is used. The PPE should also be readily
  available when working with materials that
  require it. If the proper equipment is not
  available, you should not proceed with the work.
  Notify your supervisor when you need to order
  PPE. Give an advanced notice when possible due
  to shipping time. Always consider in advance
  what you will need when conducting research.

PPE
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Maintenance of PPE

• In order to be able to rely on your equipment,
  you must take care of it. Keep the PPE clean and
  disinfected. It should be cleaned before and
  after use when possible. Proper maintenance can
  not only extend the life of the equipment, but
  also keep cost down. Keeping the cost down on
  the Personal Protective Equipment can help to
  better allocate the money toward larger equipment
  needed for research.

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What is a Sharp?
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What is a Sharp?

• A sharp is defined as any instrument, tool, or
  item that has rigid, acute edges, protuberances
  or corners capable of cutting, piercing, ripping
  or puncturing such as syringes, blades, and
  broken glass. Items that have the potential for
  shattering or breaking are also considered sharps.

http//www.ehrs.upenn.edu/training/bloodborne/bloo
dborne.html
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Safe Practices

• When using a sharp there is a risk of being cut
  by the object and possible infection occurring
  depending on what the sharp was used for. If
  hypodermic needles are used, special precautions
  must be taken to reduce the risk of a
  needlestick. After use of the needle do not
  recap, place directly in the sharp container.

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Disposal of Sharps

• All sharps must be placed into a rigid, puncture
  and leak-resistant container that is also
  impervious to moisture. The sharps container
  must be labeled either with Biohazard or
  Infectious Waste. Do not over fill the sharps
  container.

http//www.ehrs.upenn.edu/training/bloodborne/bloo
dborne.html
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Disposal of Sharps

• When the sharps container is full it must be
  collected by the EHS office. A waste pick-up
  form can be completed and a collection can be
  scheduled. A waste pick-up form can be obtained
  from the EHS office (438-8325).

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Sharps Log

• If an injury occurs as a result of a sharps
  accident, it must be recorded in the Sharps Log.
  This log is kept by the supervisor and is viewed
  upon annual inspection. The main purpose of the
  log is for risk assessment and documentation. If
  a sharps injury involves an infectious agent or
  anything beyond first aid, it is advised to seek
  medical attention at the Student Health Services.

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• What is a Risk Group (RG)?

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Risk Group

• According to National Institute of Health, there
  is a risk associated to certain research. This
  is targeting the laboratories that work with
  infectious agents and rDNA. A risk group (RG) is
  determined by the nature of the research. The
  more hazards there are with the agent/material,
  the higher the risk group and therefore more
  stringent the containment and work practices
  become.

The Risk
RG 1 RG 2 RG 3 RG 4
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Risk Groups

• The risk groups are broken down by the following

Risk Group 1 (RG1)
Agents are not associated with disease in healthy
adult humans (low individual community risk)
Example Bacillus subtilis
Risk Group 2 (RG2)
Agents are associated with human disease, which
is rarely serious, and for which preventive or
therapeutic interventions are often available
(moderate individual risk, limited community
risk) Example Streptococcus aureus
 
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Risk Groups

• Risk Group 3 (RG3)

Agents are associated with serious or lethal
human disease for which preventive or therapeutic
interventions may be available (high individual
risk but low community risk) Example Hantavirus
Risk Group 4 (RG4)
Agents are likely to cause serious or lethal
human disease for which preventive or therapeutic
interventions are not usually available (high
individual risk high community risk) Example
Ebola Virus
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Risk Groups

• The RG in your lab is to be determined by your
  supervisor. The information is to be
  communicated to you during your lab specific
  training.

Training
85
What is a Biological Safety Level (BSL)?
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Biological Safety Level

• Along with the Risk Group there is also an
  associated Biological Safety Level (BSL). This
  has the same principle as the RG in that as the
  risk increases, the containment and practices
  become more stringent. For most cases, if your
  lab is a RG 1 then the BSL is 1.
• CDC and NIH have set guidelines to help protect
  individuals who work with infectious agents
  and/or rDNA.

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Biosafety Levels (BSL)

• The different BSLs have been established as
  preventative measures against human infection.
  Taking precautionary measures can help to
  minimize hazards associated with infectious
  agents.

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BSL

• As the risk increases, the BSL also increases.
  Each BSL has specific containment practices that
  CDC and NIH have advised. When the lab specific
  training is conducted by your supervisor, she/he
  will go through the BSL guidelines for your lab.
  

Training
89

• Signs and Labeling

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Labeling

• Biological material that others may have access
  to should be labeled. This would include items
  such as cultures. The label should describe what
  the content is and also any precautions. An
  example is below.

Escherichia coli Precautions Must use Eye
protection, lab coat, gloves Only use under BSC
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Labeling

• Chemical labeling has been briefly touched on
  earlier in the presentation. One must remember
  that if any chemical is transferred to a
  secondary container, this container must be
  labeled. If the chemical will be used by the end
  of the work shift, then labeling is not
  necessary. Good science practices would
  encourage you to label all containers.

So what is included on the Label? Click here
92
Signs

• Signs are a way of communicating important
  information. It is a way to heighten awareness
  about hazards that exist.
• There may be signs on laboratory refrigerators
  reminding you that no food or drink can be stored
  in it. There may be radioactive or biological
  materials that could possibly be absorbed in
  food.
• There may also be signs which denote that PPE
  must be used.

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Signs

• Labs which use rDNA and infectious agents must
  have a sign posted on the outside of the door.
  Before someone enters the lab, they will have the
  information they need to protect themselves.
  Always read the signs carefully so you know what
  precautions to take.

94
Containment of Laboratory
95
Containment
Containment of a laboratory is to confine
organisms containing rDNA molecules and/or
infectious materials. The purpose is to reduce or
eliminate the potential for exposure of the
laboratory worker, persons outside of the
laboratory, and the environment from organisms
containing rDNA and infectious materials.
96
Containment
There are three elements to containment
Laboratory Practices Containment
Equipment Special laboratory design
Lets take a closer look
97
Practices and Techniques
Responsible practices and techniques are required
when working with hazardous materials. This is
one very important way to reduce
accidents/injuries. The following practices
should be followed in addition to reviewing the
Biosafety Manual
Practices and Techniques
98
Safety Equipment

• The Primary Barriers

99
Safety Equipment
Certain equipment is necessary to achieve
compliance and most importantly to provide
adequate protection. The safety equipment that
is needed is known as primary and secondary
barriers. Lets take a look at the difference in
the barriers.
100
Primary Barriers

• Primary barriers are referring to protective
  measures including engineering controls. This
  includes not only PPE that has already been
  covered, but it also includes safety cabinets,
  fume hoods, vaccines and autoclaves.

It is important to know when this equipment is to
be used and how to properly use it.
101
Primary Barriers

• Lets look at the following barriers
• (Besides PPE)
• Fume hood
• Biological Safety Cabinet (BSC)
• Autoclave

102
Fume Hood

• The fume hood is used with chemicals. The main
  function is to exhaust the vapors and gases that
  are generated in the hood to the outside. The
  hood is designed to minimize your exposure to
  airborne contaminants. This is not to be used
  with biohazardous materials.

103
Fume Hood Use

• When using the fume hood you first need to make
  sure the exhaust blower is operating and air is
  entering the hood.
• Remember, do not put your face inside the hood!
• Minimize storage of chemicals in the hood
• Clean spills immediately
• Work with the sash at the proper operating level
  as indicated by the arrows

104
Biological Safety Cabinet

• The biological safety cabinet (BSC) is used as a
  containment for infectious agents. The BSC has a
  HEPA filter in the exhaust system to protect the
  environment and yourself.
• The (HEPA) filter is a high efficiency
  particulate air filter. It is able to remove
  particles at a size of 0.3 ?m with an efficiency
  of 99.97. It is also able to remove both
  smaller and larger particles.

105
BSC

• There are 3 classes of BSC that are used. The
  higher the risk group and biosafety level, the
  higher the class of cabinet that is used.
• If there is an infectious agent being used,
  whether it is used in research animals or
  cultured, it must be manipulated inside the BSC.

106
BSC

• When using this containment device, remember to
  also use the proper personal protective
  equipment. The following PPE should be
  considered depending on the BSL that is required
  for the organism that you use.

Gloves Lab Coat Shoe Covers Safety
Glasses Respirator
107
Use of BSC

• Always make sure that the BSC has been
  decontaminated both before and after use.
  Decontamination methods vary depending on the
  infectious agent being used. Once the
  decontamination of the cabinet is complete, place
  the waste in a biohazard bag for autoclave.

108
Use of BSC

• Do not bring contaminated materials out of the
  cabinet until they have been surface
  decontaminated. If you are unable to
  decontaminate, place the material into a closed
  container to transfer it to the autoclave

109
Work Practices for the BSC

• Remember to follow the work practices
• Work in such a way that your face is above the
  front opening
• Wait for 1 minute after placing hands/arms
  inside the cabinet to stabilize the air flow
• Work at least 4 inches from the inside edge of
  the front of the grille

110
Work Practices for the BSC

• Remember to place all materials as far back in
  the cabinet as possible
• Limit the storage within the cabinet

111
Autoclave

• An autoclave is used to treat infectious
  material and rDNA. As a standard for the
  University, all material of this nature must be
  autoclaved as a safety precaution. The autoclave
  is able to render the material as non-infectious.

112
Autoclave

• The autoclave is able to reach a high
  temperature to sterilize the agent. It is
  important to know the standard operating
  procedures (SOP) for the autoclave. The SOP is
  located next to each autoclave. If the
  temperature or pressure is inadequate, the bag is
  overfilled, or the peak time is not long enough
  the material will not be properly decontaminated.
  

113
Autoclave

• The autoclave is sensitive to time, temperature,
  pressure, water content, type of container being
  used and the type of waste being sterilized.
  When using the autoclave there are a few
  important point to remember in order for the
  process to be effective.

114
Procedures

• The material being autoclaved must be placed
  inside an autoclave red bag or in a pan (Do not
  overfill the bag).
• Add 250-500ml water to the bag or pan for steam
  to generate and properly circulate
• A spore strip or other approved tape must be
  placed on the outside of the bag or pan to verify
  that the heat treatment was successful. You will
  have to verify that a color change took place.

115
Procedures

• The autoclave must reach a temperature of 121C
  (250 F) for at least ½ hour at a pressure of
  15psi
• After successfully being autoclaved, the
  material can be placed in regular trash.
• Sharps must be incinerated. When the container
  is full, a pick up can be scheduled through EHS
  and from there Student Health Services will
  incinerate the sharps container for proper
  disposal

116
Autoclave Log

• Each time the autoclave is used, the log must be
  completed by the individual using it. The log is
  to be located at the site of the autoclave. The
  information contained within the log is as
  follows

• Date
• Generator Name (P.I.)
• Location
• Time the autoclave reached 121C
• Max Temp.
• Pressure
• Type of waste
• If spores or tape were used
• Weight
• Person using autoclave

117
Autoclave Log

• The log is required by EPA and it must be
  maintained for 3 years by the department.
• The purpose of the log is to ensure that proper
  decontamination is taking place
• If a new log is needed at the autoclave, contact
  biology department or EHS.

118
Facility Design

• The Secondary Barriers

119
Secondary Barriers

• Secondary barriers are the facility design and
  construction. These barriers are to provide
  protection for the individuals outside the lab,
  the community, and the environment.

120
Secondary Barriers

• An example of a Secondary barrier would be a
  laboratory. It is separate from the classroom
  and has limited access.
• The animal room has limited access and is only
  available to authorized personnel.
• The autoclave is set away from the normal
  traffic.
• The handwashing facilities are located within
• the lab and not accessible by the outside
  students.

121
Decontamination
122
Decontamination

• Decontamination is the removal or neutralization
  of toxic agents or the use of physical or
  chemical means to remove, inactivate, or destroy
  living organisms. This includes both
  sterilization and disinfection.

123
Decontamination

• Decontamination is the responsibility of all
  laboratory workers. Failure to decontaminate can
  result in exposure to infectious agents which can
  cause great illness. Most decontamination can be
  done by chemicals. This technique is used only
  when autoclaving is not possible. Continue on to
  see what would be best for your lab.

124
Chemicals

• There are a variety of chemicals that can be
  used as an effective method of decontamination.
  Depending on the agent being used, the method to
  use may vary along with the contact time. For
  most organisms, a 1100 chlorine solution for
  10-30 minutes is adequate. The Biosafety Manual
  has a list of sterilizers/ disinfectants that can
  be used.

125
Autoclave

• The autoclave is the most effective method to
  use for decontamination purposes. As a general
  rule of thumb, autoclave all materials that are
  considered infectious agent, rDNA, or resemble
  components of this nature. When in doubt,
  AUTOCLAVE! If a material is not capable of
  autoclave because of its size, material, or it is
  stationary, then rely on chemical disinfectant as
  a second option.

126
Incineration

• There are other methods that can be used for
  decontamination such as incineration. This is
  the most reliable method, but it will destroy all
  materials placed in it and it creates
  environmental pollutants. If you must
  incinerate, contact EHS.

127
Spills and Accidents
128
SOP

• Spills and accidents can pose a serious health
  and safety threat. When a spill occurs, an
  aerosol can be created which can make the
  material several times more potent. The best
  measure to take in order to protect yourself is
  to be prepared. There should be standard
  operating procedures for this type of situation
  in your lab.

129
What to Do?

• Being able to recognize the hazards, mitigate
  the spill, and notifying response authorities can
  be your best defense. The first response to a
  spill should be to evacuate the immediate area
  until the scope of the hazard has been addressed.
  Seek medical attention if necessary. Allow
  sufficient time for the aerosol to settle before
  considering entering the room. If you are
  responsible for clean up, proper training shall
  be addressed.

130
How to Report

• When a spill occurs, it must be recorded in the
  Spill Log. The main purpose of the log is to
  track an exposure in case of future
  illness/injury. Report to your supervisor all
  spills. If medical attention is needed, it is
  suggested to go to Student Health Services.

131
Waste Management
132
Waste

• Hazardous and biohazardous waste has special
  guidelines for proper disposal. It is important
  to properly dispose of waste to ensure human and
  environmental health. EPA regulates the waste
  that is generated at ISU.

133
Waste

• Waste can be classified as either hazardous or
  biohazardous. Lets take a closer look at the
  differences.
• Hazardous Waste- This is a waste which contains
  the characteristics of being any of the
  following

Toxic Corrosive Ignitable Flammable Oxidizer
134
Waste

• A biohazardous waste is any waste that is
  considered infectious and/or because of its
  biological nature it can cause physical or health
  hazards in humans, animals, plants or the
  environment. This includes recombinant DNA and
  other genetically altered organisms and agents.

135
Proper Disposal

• Waste that is considered biohazardous can be
  disposed of in regular trash once it has been
  rendered non-infectious. If a biohazard labeled
  bag is used, make sure it is either placed in a
  secondary bag or a completely new bag that is not
  red.

http//www.lbl.gov/ehs/Medical/html/biohazardous.h
tm
136
Proper Disposal

• Hazardous waste must be disposed of through the
  Environmental Health Safety office. If you
  need to have hazardous waste picked up, contact
  EHS for a pick-up form.

EHS 438-8325
137

• You have now completed the General Lab Safety
  Training presentation. If you have any questions
  please take the time to ask your supervisor or
  call EHS. By signing the training form, you are
  stating that you have completed the training and
  that you understand the content within the
  training.

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