General Laboratory Safety Training Developed for Science Laboratories September 2002
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Developed for Science Laboratories
There are hyperlinks throughout the presentation. Where there is the following icon additional training is required.
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.
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.
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.
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.
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.
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.
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.
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.
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!
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.
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. Let’s take a look.
A health hazard has the following characteristics:
A physical hazard has the following characteristics:
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.
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.
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.
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.
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. Let’s look at these hazards a little closer.
A biological hazardincludes 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.
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.
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.
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.
Biosafety levels will be discussed in detail by your supervisor during the Lab Specific Training.
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.
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.
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.
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.
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.
You should remember the following:
Personal habits play a large role in minimizing hazards. The following measures must be taken:
These safe practices should be followed to ensure safe working conditions:
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:
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.
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.
According to the laboratory standard, for those labs that have chemicals the following training topics must be addressed:
This training is covered in the Hazard Communication Training Presentation
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.
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.
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.
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.
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:
There are several constituents that are covered in the Laboratory Standard. Among these constituents includes labeling and the MSDS. Let’s look more closely at the content of the 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.
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!
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 Biological Agent
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.
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.
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.
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.
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.
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.
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!
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!
When chemicals are being used there is always the possibility of splashing. The proper PPE to use when chemicals are involved would include:
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.
When biological agents and/or rDNA are being used the following PPE must be used:
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.
When animals are being used the following PPE must be used:
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.
Depending on the arthropod that you encounter in the lab, the PPE could vary. In most cases the PPE below should be used used.
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.
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.
If you think you need a respirator talk with your supervisor or contact EHS (438-8325)
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.
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.
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.
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.
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.
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).
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.
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 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
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
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.
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.
The different BSLs have been established as preventative measures against human infection. Taking precautionary measures can help to minimize hazards associated with infectious agents.
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.
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.
Must use Eye protection, lab coat, gloves
Only use under BSC
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
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.
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.
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.
There are three elements to containment:
Special laboratory design
Let’s take a closer look
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
The Primary Barriers
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. Let’s take a look at the difference in the 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.
Let’s look at the following barriers:
Biological Safety Cabinet (BSC)
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.
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
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.
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.
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.
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.
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
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
Remember to place all materials as far back in the cabinet as possible
Limit the storage within the cabinet
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.
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.
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.
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.
The autoclave must reach a temperature of 121°C (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
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:
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.
The 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.
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.
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.
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.
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 1:100 chlorine solution for 10-30 minutes is adequate. The Biosafety Manual has a list of sterilizers/ disinfectants that can be used.
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.
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.
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.
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.
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.
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.
Waste can be classified as either hazardous or biohazardous. Let’s take a closer look at the differences.
Hazardous Waste- This is a waste which contains the characteristics of being any of the following:
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.
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.
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.
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