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Purpose Chemical Emergency Procedures Objectives Biological Safety in Laboratories Safety Responsibilities Field Safety Guidelines Standard Operating Procedures Training Chemical Safety in Laboratories Medical Evaluations Electrical Laboratory Equipment Forms: Special Provisions for Particularly Hazardous Materials Student Permission to Work Independently After Hours Form Project Hazard Assessment Form Student Visitor Accident Report Form Memorandum of Understanding and Agreement Registration of Biohazardous Materials and Recombinant DNA Guide to the Globally Harmonized System of Classification and Labeling of Chemicals (GHS)
Protecting the health and safety of every person on the University of Houston-Downtown (UHD) is of paramount importance, and it is a continuous work in progress. The UHD Laboratory Safety Manual (LSM) constitutes written standard operating procedures designed to ensure laboratory operations are carried out in a manner that protects all persons from harmful exposures associated with hazardous chemicals, microbial, biological and general laboratory hazards, as well as, complying with state and federal regulations.
General and Chemical Safety sections of this manual were taken from other safety manuals available online including one from TAMU – Arkansas. Biological Safety sections were taken from the published Biosafety in Microbiological and Biomedical Laboratories guidelines from the www.cdc.gov. Much of the information in the field safety section came from “Safety Guidelines for Field Research,” Office of Environment, Health & Safety, University of California, Berkeley.
The UHD LSM shall be used in all current and future laboratories. The manual will be continuously reviewed and updated as needed and/or required. The safety and well-being of all persons inside UHD laboratories will be accomplished by our dedication to achieving the following goals:
- Provide the necessary facilities, staff and equipment to operate in the safest manner possible.
- Assure that staff, faculty and students receive appropriate training for the experiments they are engaged in.
- Reduce or minimize the extent of chemical, microbial, biological and hazardous exposure by all laboratory personnel.
- Provide adequate ventilation to all laboratories.
- Observe the threshold limit values for all chemicals.
- Ensure the chemical inventory in labs is constantly maintained in a manner that ensures only the least amount of required chemicals are on hand for the specific needs of the department, as well as, constant monitoring of the inventory to dispose of chemicals that have reached the end of their shelf life.
- Protect the environment from hazardous chemicals, chemical waste, microbial contamination and biohazards.
- Assure that all risks are properly assessed, and ensure that all experiments with microorganisms or biohazards comply with federal safety guidelines.
- Establish guidelines for safe data collection in the field.
- Ensure all of the above goals are being met by inspecting all laboratories on a periodic basis
Responsibility for Safety in the laboratory rests with the Chief Safety Officer (CSO), the Principal Investigator (PI), and the laboratory worker. The UHD President has the ultimate responsibility for the Laboratory Safety program within the University and must provide continuing support for institutional laboratory safety.
The Chair of the Department of Natural Sciences shall be the designated Chief Safety Officer (CSO) for all experiments in Natural Sciences. The NS CSO shall provide guidance to the Principal Investigator (PI) on developing and implementing safety plans and for providing technical assistance in developing standard operating procedures related to laboratory safety and emergency response. The CSO is responsible for giving prior approval for any operation which presents a foreseeable hazard to any person.
The CSO shall designate all faculty involved in research as Principle Investigators (PIs). All PIs shall be responsible for developing and implementing a Laboratory Safety Plan for their experiments. The PI may use the guidelines in this LSM as the basis for their Laboratory Safety Plan. Supplemental safety information must be added if not covered in this manual. The PI is responsible for the health and safety of all personnel under his/her direction. Specific responsibilities include:
- Reviewing the LSM on an annual basis, ensuring it is continuously updated as to current chemical safety procedures.
- Monitoring the purchase, use and disposal of chemicals and biological waste used in laboratory procedures.
- Ensuring all lab personnel and students are properly trained in the use of hazardous chemicals and biohazards.
- Maintaining documentation of training.
- Ensuring all personnel and students know and follow the rules of laboratory safety.
- Ensuring engineering controls are operative and personal protective equipment (PPE) is properly selected, used and maintained.
- Reviewing specific hazards for each new chemical or biological introduced into the lab and training personnel/students in safe handling of new materials.
- Ensuring Material Safety Data Sheets (MSDS) are available or immediately accessible online for each hazardous chemical found in the laboratory.
- Filing written reports with the CSO and Environmental Health and Safety (EHS) Office concerning chemical spills, release of biohazards and any accident or injury that occurs in the laboratory. (Paperwork should be initiated in the NS office.)
- Ensuring regular laboratory safety and housekeeping inspections are performed.
- Understanding and keeping current of all legal requirements concerning hazardous chemicals and biological hazards.
- Ensuring laboratory equipment is properly maintained and in good working order.
All laboratory employees and students are responsible for:
- Knowing and following the proper safety procedures in the LSM.
- Reporting all hazardous conditions to the PI.
- Wearing the prescribed personal protective equipment (PPE).
- Reporting any injury, illness or chemical/biological spill to the PI and/or the NS Office.
- Requesting information or training when unsure of how to handle a hazardous chemicals or biological hazards.
This portion of the LSM represents an initial minimum set of guidelines for experimentation in UHD laboratories.
Every PI, staff member and student must be aware of the following general guidelines.
- Students working in laboratories must be under the supervision of a PI and only UHD employees, students and approved visitors are allowed in UHD laboratories.
- The safety guidelines must always be applied to any experiments being performed.
- There should be training on the location and the appropriate use of fire extinguishers, eyewash stations, emergency showers, fire exits, evacuation routes and fire alarm pull stations.
- There should be training on the types of personal protective equipment (PPE) available and how to use it appropriately.
- Equipment must be in good working order before conducting and experiment (no error messages of equipment, glassware is not chipped or broken, etc.)
- Equipment can only be used if the experimenter has appropriate training and may only be used as directed.
- Experiments that can be left safely unattended for any length of time must be properly identified with a posted sign that notifies the reader of the contact person, cell phone number, and any hazards or considerations of the experiment.
- Hazardous materials will be used with caution and only with proof of proper training. Very hazardous materials, such as carcinogens, reproductive toxins and chemicals with a high degree of acute toxicity, will not be purchased or used without prior written approval from the CSO. The CSO must weigh the need for conducting operations using these type chemicals with the degree of risk of exposure to staff and students. If the risk outweighs the need, the CSO shall deny the request. Students may be denied approval to work hazardous materials by the CSO. Hazardous materials can only be used for their intended purpose.
- If an injury occurs in the lab that requires medical attention, call 911 or x8911 immediately. All injuries or near misses must be reported to the NS office (N-813) immediately. A report of Safety/Health Hazard and Near Miss form will be completed. Students and visitors will also have to complete a Student Visitor Accident Report Form.
- If exposed to a toxic/hazardous material, immediately consult and enact decontamination procedures as outlined in the appropriate MSDS or other safety document.
Working in laboratories from 7:00AM to 7:00PM Monday through Saturday is defined as working “during regular hours.” Working in laboratories from 7:00 PM to 7:00 AM is defined as working “after hours.” Undergraduate research students or students in courses which require after-hours work will need to have their faculty member complete the Student Permission to Work Independently After Hours Form . This must be approved prior to beginning work.
Policies for Student Independent Research Work in the Laboratory
A student may use laboratory facilities for independent research only after the PI in charge has reviewed the experimental procedure for any associated hazards and has determined that the student possesses adequate training in proper experimental and emergency procedures. Students will be given a copy of the Project Hazard Assessment form (PHA) filled out by his/her PI which describes the procedures and hazards associated with working in this particular lab. Students must sign the PHA and are responsible for performing all their work in accordance with those procedures. Students must report all accidents, chemical spills, and unsafe conditions to the PI who is responsible for making sure proper paperwork is completed as necessary. Students working with chemicals or biohazards must also satisfactorily complete safety training.
Students must have a completed and approved Student Permission to Work Independently After Hours Form prior to working after hours in a laboratory (Appendix A). The permission form must be completed and on file in the NS office. If the after-hours work involves the use of chemicals or biological hazards, the permission form must also indicate that safety training has been successfully completed. The CSO is ultimately responsible for any laboratory activity which presents a foreseeable hazard to employees, students or structures. As such, the CSO may not grant approval for after-hour student work that involves a hazardous or potentially dangerous situation.
Policies for Students Enrolled in Laboratory Courses
Students enrolled in laboratory sections of a course are allowed to perform laboratory procedures alone only when the PI or instructor authorizes that this is acceptable. When working in the laboratory during regular hours, students must be under the direction of a qualified person – a PI or course instructor who is familiar with emergency procedures. Properly directed students can work in a laboratory course outside of the regular laboratory periods (but during regular hours) only with the permission of the instructor. The instructor is responsible for ensuring that the students receive sufficient instruction to work safely under these circumstances. Students enrolled in a laboratory course may not work after-hours unless authorized by the CSO with an accompanying Permission to Work Independently After Hours form on file in the NS office.
Emergency eyewash stations are located in the UHD chemical and microbiological laboratories on the 8th and 9th floors of One Main Building only. (There are surge hoses that will be used as a safety shower for other laboratories.) Eyewash stations should be kept free of any obstruction which might inhibit their use. Eyewash stations should be tested on a monthly basis to verify the units are in proper working order and to flush the lines of stale water and debris. The EH&S office is responsible for ensuring these stations are checked. Documentation of the time, date and the person’s name that performed the check shall be kept.
Fire Extinguishers and First Aid Kits
Each laboratory will be equipped with fire extinguishers and first aid kits. Everyone working in a laboratory should know how to properly use the extinguisher. Fire extinguishers shall be mounted on an interior lab wall in a highly visible and accessible area. Each extinguisher will be checked on an annual basis to ensure it is in good working order. First aid kits must be checked by the PI during regular equipment checks.
Personal Protective Equipment (PPE) is any item designed to protect laboratory personnel from exposures and injury. The PI is responsible for the selection, procurement, care and replacement of all personal protective equipment. PPE should never be worn outside of the laboratory.
Closed-toe shoes must be worn in all laboratories. The PI will instruct all laboratory personnel on the use of all mandatory PPE for use in a particular laboratory and for particular experiments. It is recommended that employees and students wear clothing that minimizes exposed skin surfaces. When a lab coat is used, it should be worn over street clothes. Lab coats are intended to prevent contact with dirt, chemical dusts and minor chemical splashes or spills. If they become contaminated, they should immediately be removed and the affected skin surface washed thoroughly. Sandals or open-toed shoes may not be worn in laboratories. Long hair must be tied-back.
Eye protection is required for all personnel and any visitors present in locations where chemical splash hazards exist. Safety glasses, goggles and full face shields are the three main types of eye protection. Safety goggles should be worn in situations where bulk quantities of chemicals are being handled and chemical splash to the face is possible. Goggles and full face shields should be worn when handling highly reactive substances or large quantities of hazardous chemicals, corrosives, poisons and hot chemicals. All eye protection should meet the requirements of the American National Standards Institute (ANSI) standard, Z87-1. Material Safety Data Sheets (MSDS) will list the recommended PPE for use with each chemical.
When inserting tubing into stoppers, lubricate tubing and wear gloves to protect hands from being cut in the event of the tubing breaking.
Gloves are essential when working with hazardous substances. The proper gloves will prevent skin absorption, infection or burns. Chemical resistant gloves should be worn whenever the potential for contact with corrosive or toxic substances and substances of unknown toxicity exist. Glove selection should depend on the type chemical being handled, the particular hazard involved, and the glove’s suitability for the operation being conducted. Before each use, gloves should be checked for integrity. Non-disposable gloves should be replaced periodically, depending on frequency of use and their resistance to the substances being handled. Disposable gloves should be worn when working with biohazards. All gloves should be removed prior to touching keyboards, phones, door knobs and other surfaces and should not be worn outside of the laboratory. After glove removal, hands should be washed before leaving the laboratory.
Inhalation hazards can be controlled using ventilation or respiratory protection. Check the chemical label and MSDS for information on a substance’s inhalation hazard and special ventilation requirements. When a potential inhalation hazard exists, the label or MSDS will contain special warnings. Take appropriate precautions when handling these substances. Controlling inhalation exposures through engineering controls (ventilation) is always the preferred method. As with other PPE, respiratory protection relies heavily on employee work practices and training to be effective. UHD does not use any chemicals at this time requiring the use of a respirator. In the future, if it becomes necessary to use chemicals requiring respirators, a Respiratory Protection Program will need to be put in place before the chemical can be introduced into the lab or any other area of the University.
Professional behavior is expected at all times in the laboratory. No unauthorized persons, including children, shall be allowed in the laboratory. No food or drinks are ever allowed in the laboratory. All laboratory work areas must be kept clean and clear of obstructions and clutter. Floors must be kept dry and aisles shall remain clear of boxes or other items that could be trip hazards. All laboratory wastes shall be kept in appropriate containers and labeled accordingly. If large amounts of trash require disposal, facilities management should be contacted immediately. Violations will not be tolerated and will result in the removal of the offender from the laboratory.
MSDS sheets should be the first source of information about the hazards associated with any chemical. Manufacturers are required to provide a MSDS for each chemical product they sell. All laboratory personnel will have access to MSDS sheets in their laboratory via the internet or hardcopy. A backup copy of all MSDS sheets will be housed in the chemical stock room.
MSDS sheets should contain the following information:
- Name, address and phone number of the manufacturer
- Chemical name, synonyms, and Chemical Abstract Number (CAS)
- Physical properties
- Health hazard information (most MSDSs use NFPA or HMIS hazard ratings)
- First aid measures
- Firefighting measures
- Handling and storage precautions
- Exposure controls/personal protection
- Stability and reactivity
Most MSDS sheets will also contain the following information:
- Toxicological information
- Ecological information
- Disposal consideration
- Transport information
- Regulatory information
Before any task involving hazardous materials or physical hazards is performed, the PI must have completed the PHA for the project and it must have been approved by the NS Safety Committee and have all signatures (PI, NS Safety Committee Representative, Chair, Dean and EH&S Manager). The PHA should include Standard Operating Procedures (SOP) for dealing with any listed hazards. The NS Safety Committee, Chair, Dean or EH&S Manager may reject a PHA if it is determined that it has not been completed sufficiently or if it is judged that the experiment is too hazardous to be completed at UHD. The PHA should not be limited to chemical hazards, but should also include biological hazards, heat and cold hazards, physical hazards and hazards exposed to in the field.
The PI for any laboratory shall be responsible for the procurement of all lab chemicals. Employees involved in the receiving of chemicals should be informed about proper handling, storage and disposal procedures. Chemicals should not be accepted without accompanying labels, MSDS and proper packaging. Damaged or leaking containers shall not be accepted for any reason. All chemicals should be dated upon receipt. The PI should always inform employees and students about the proper handling of any new chemicals introduced into the laboratory.
The laboratory PI is responsible for the instruction of all laboratory personnel in the safe use of chemicals. Information regarding the classification and safe use of chemicals can be found at the following web site: http://www.osha.gov/dsg/hazcom/ghs.html.
Carts used for transport should be sturdy and have a substantial rim around the edge. All chemicals should be transported in an appropriate secondary containment vessel.
The following is a list of important safety rules for the storage of chemicals:
- Store chemicals according to their chemical classification.
- Never store chemicals anywhere other than an approved storage area.
- Store especially hazardous chemicals in a secondary container.
- Never store chemicals in a fume hood.
- Return chemicals to their storage areas at the end of the day
- Store flammable chemicals that should be refrigerated in an approved, explosion-resistant refrigerator that is labeled as such.
- Never stack bottles on top of each other.
- Label all chemical containers with the date of receipt and the date opened.
- Be aware of procedures for transferring chemicals that may be needed for proper dilution or storage. (For example, never add water to concentrated acid, etc.)
The security of the laboratory chemical inventory rests with the lab PI. All PIs are required to inventory their chemicals when they initially set up their labs. Subsequent shipments of chemicals must be dated and the PI shall also conduct an annual inventory of all chemicals on hand. A copy of this inventory shall be maintained in the laboratory and a copy shall be sent to the chemical stock room, the NS office and then to the EH&S office. Additionally, the following principles should be applied in an effort to keep an accurate track of laboratory chemicals.
- Before any work involving hazardous chemicals is performed, turn the hood fan on and make sure it is working.
- Chemicals must be purchased in limited amounts. A (6) six-month supply is preferred.
- An expiration date should be assigned to each chemical container. Expired chemicals will be properly disposed of.
- Chemicals should be examined annually. During this inspection, chemicals which are beyond their shelf life, show deterioration of the chemical, have questionable labels, are in leaking containers or have corroded caps will be eliminated.
All hazardous laboratory work should be carried out in an appropriate chemical fume hood. All chemical fume hoods should be regularly inspected and certified on an annual basis. Uncertified hoods should not be used.
- Before any work involving hazardous chemicals is performed, turn the hood fan on and make sure it is working.
- Check the inspection sticker to make sure the hood has been inspected within the last year.
- Fume hoods should be used with the sash positioned at 18 inches or less when possible for optimal performance.
- All equipment should be kept at least 6 inches inside the face of the hood to prevent disruptive airflow patterns.
- No chemicals should be stored in fume hoods (either for use or for disposal).
- Maintain an air space under large equipment by placing on blocks to allow air currents to freely pass under the equipment. This minimizes “dead space” at the hood face and thereby improves overall hood performance.
- Do not use the fume hood as a storage cabinet. Excessive storage can obstruct airflow and cause areas of low air velocity at the face opening.
- Do not put your face or head inside the hood.
- Minimize sources of cross drafts (open windows, doors, fans, etc.) which may pull contaminated air from the hood.
- Ensure all fume hood users are aware of safety procedures in case of an emergency.
Hazardous chemicals used in the UHD laboratory will be disposed of in a safe, approved manner consistent with all applicable laws and the UHD Campus Waste Management Program. Laboratory staff should use the smallest quantity of hazardous substance that is practical in an effort to generate the least amount of hazardous waste. This should include reviewing experimental protocols with the goal of substituting non-hazardous or less hazardous reagents, using micro-scale procedures, and using materials that can be easily neutralized or detoxified. UHD has an approved, underground chemical neutralization/dilution system that is connected to the laboratory sinks. Small quantities of lab chemicals can be flushed down the sinks. Maintenance of the dilution tank as well as the approved disposal of any leftover chemical waste in the tank is the responsibility of the Director of the Physical Plant.
The utilization of electrically powered equipment can pose hazards in the laboratory when not used properly. Problems that are encountered when using any lab equipment should be reported to the laboratory supervisor immediately.
General Electrical Safety
The use of an autoclave is the preferred way to decontaminate infectious waste. In order to safely operate the autoclave, the following procedures must be utilized:
- Do not put sharp or pointed contaminated objects into an autoclave bag. Place them in an appropriate rigid sharps container.
- Never use red biohazard bags to autoclave.
- Always use caution when handling an infectious waste autoclave bag in case sharp objects were inadvertently placed in the bag. Never lift a bag from the bottom to load it into the chamber, lift from the top of the bag.
- Use indicator tape with each load to verify it has been autoclaved.
- Do not mix contaminated and clean items together during the same autoclave cycle.
- Be sure to wear personal protective equipment, including heat resistant gloves, safety glasses and a lab coat when operating an autoclave. Caution should be used when opening the autoclave door, allowing superheated steam to exit before removing the contents.
- Always be on the alert when handling pressurized containers as superheated liquids may spurt from closed containers. Never seal a liquid container with a cork or stopper as this could cause an explosion inside the autoclave.
- Use caution when autoclaving agar or trash with agar as it will become liquefied when autoclaved. Solid autoclave buckets (not baskets) should always be used with agar.
- If there is a spill inside the autoclave chamber, allow the unit to cool before attempting to clean up the spill. If glass breaks in the autoclave, use tongs, forceps, or other mechanical means to recover fragments. Do not use bare or gloved hands to pick up broken glassware. Do not attempt to fix or clean the autoclave without help of a PI or Instrument Technician.
All centrifuge operators shall be instructed on proper operating procedures, which would include balancing loads, selection of proper rotor, head, cups, and tubes, and use of accessory equipment. (Consult the centrifuge operating manual.) Additionally, the following guidelines should be followed when dealing with centrifuges.
- Students may not be use a centrifuge until they have been properly trained. Operating procedures for each centrifuge must be established by the laboratory PI in accordance with the procedural outlines in the operating manual. Guidelines for centrifugation of infectious agents or chemical hazards must be included in the procedures.
- The centrifuge operator is responsible for the condition of the machine at the end of each procedure.
- All centrifuge tubes must be inspected before each use. Broken, cracked or damaged tubes should be disposed of.
- Refer to the centrifuge operating manual for selection of appropriate tubes, carrier cups, and rotors. Capped centrifuges should be used whenever possible.
There should not be any potential sources of electrical sparks on the inside of a laboratory refrigerator where chemicals are to be stored.
- If the unit will be used to store flammable or combustible materials, the refrigerator/freezer should be explosion-resistant and labeled to indicate it is suitable for storing flammable materials.
- The explosion-proof refrigerator should be used to store flammable or combustible materials in hazardous areas, such as a chemical storage rooms.
- All refrigerators, regardless of the type, should never be used to store food and must have labels on the door indicating this.
Drying ovens are commonly used to dry laboratory glassware. Since these ovens do not have a provision for preventing the discharge of volatized substances into the air, organic compounds should not be dried in these units. Conventional oven units should not be used to dry any chemical that is moderately volatile and might pose a health hazard of acute or chronic toxicity. Glassware rinsed in organic solvent should not be dried in an oven.
In addition to general safety guidelines, special precautions are needed when handling select carcinogens, reproductive toxins, and substances that have a high degree of acute toxicity. The PI must ensure that the below listed precautions are taken in order to minimize risk of exposure to these substances. Again, a Project Hazard Assessment Form be filled out and approved before any project with hazardous materials may be considered.
- Quantities of these substances used and stored in the laboratory should be minimized, as should their concentrations in solutions or mixtures.
- Work with genotoxins (chemicals that damage the genetic material of humans), reproductive toxins, and acutely toxic chemicals should be performed within a functioning hood, biological safety cabinet, sealed system, or other system designed to minimize exposure to these substances. (The exhaust air from the ventilation system may require scrubbing before being released into the atmosphere.) In all cases, work with these chemicals must be done in such a manner that the OSHA permissible exposure limits (PEL) or similar standards are not exceeded.
- Compressed gas cylinders that contain acutely toxic chemicals, such as arsine and nitrogen dioxide, should be kept in ventilated gas cabinets. The ventilation efficiency of the designated ventilation hood or gas cabinet and the operational effectiveness of equipment used to contain or manipulate these special substances should be evaluated periodically by the NS Equipment Technician or EH&S personnel. The interval of evaluating systems may vary from weekly to semi-annually depending on the frequency of usage, the quantities employed, and the level of the hazard.
- UHD laboratories utilizing hazardous substances must have designated areas for this purpose, marked with the appropriate hazard signage. The designated area may be the entire laboratory, an area of the lab, or a device such as a ventilation hood or glove box. The designated area must be marked with a DANGER, specific agent, AUTHORIZED PERSONNEL ONLY, or comparable warning sign.
- All faculty, staff or students who work in a UHD lab which has an area designated for use with hazardous chemicals must be trained regarding the serious harmful effects of these substances as well as the signs and symptoms of exposure to these substances, whether or not they actually work with the substances themselves.
- UHD faculty, staff or students working with these substances must have access to the appropriate personal protective equipment and clothing, and must be trained on how to properly utilize the safety equipment.
- Detection equipment will be required if chemicals (especially poisonous gases) with a high degree of acute toxicity are utilized.
- All waste contaminated with these substances should be collected and disposed of in a timely manner as outlined in the UHD Hazardous Waste Program.
- The designated work area shall be thoroughly decontaminated and cleaned at regular intervals.
- Special precautions to avoid release and exposure to highly toxic chemicals, reproductive toxins and genotoxins must be utilized. For instance, volatile substances should be kept cool and contained. Gases should have properly functioning valves, check valves and regulators. Containers that can withstand pressure buildup, and the appropriate piping, should be kept in closed areas with minimal air currents. Appropriate contact materials should be used to avoid static charging.
- Emergency response planning for releases or spills must be prepared by the PI, approved by the CSO and included in the training of staff, students and all others that may be affected in the building.
All laboratory personnel need to know what to do in the event of a chemical spill or other emergency that may occur in UHD laboratories. Most spills can be avoided if proper spill prevention guidelines are followed. While most spills can be safely cleaned up by those who caused the spill, some spills should only be cleaned up by specifically trained emergency response personnel. The CSO, PI, and the Health & Safety Officer must be notified whenever there is a chemical spill that poses any health or environmental risk. An analysis of the spill will be conducted in order to find out the cause of the spills and what methods should be taken to prevent it from happening again.
To prevent spills labs should reduce clutter and unnecessary materials in the work areas, eliminate tripping hazards and collect all of the needed equipment before starting work. When storing chemicals and supplies they should be stored on sturdy shelves (shelves with lips are preferred) with larger containers stored close to the floor. Containers should be pushed to the back of shelves and unprotected glass containers should never be stored on the floor.
If chemicals must be transferred from one area to another, then carts, safety containers, bottle carriers should be utilized as appropriate. All potential hazards should be considered before transporting chemicals so that spills can be prevented.
Precautions for Transferring between Containers
If transferring chemicals it is necessary to pay careful attention to the size of the container being used to avoid overfilling. Pumps or other mechanical devices should be utilized when possible rather than simply pouring a chemical. When transferring it may also be necessary to pour in a hood or in a secondary container to capture leaks and spills.
Materials Available for Spill
Before any hazardous chemical is used in the laboratory, the PI has the responsibility to perform a hazard assessment of the procedure to be undertaken. This would include determining what could go wrong and what to do in the event of a spill or other emergency. When performing this task, the PI should determine what PPE is needed, as well as any other safety equipment and containment/clean-up materials and to have these items on hand. Spill control kits should be available and their locations clearly marked. All staff and students should be aware of the kits, how to use them and their limitations. Suggested items for a Small Spill Kit are:
- Safety goggles
- Lab coat
- Small bag of absorbent (kitty litter)
- 5 gallon plastic bucket
- Heavy gloves appropriate for the material
- Acid/Base neutralization materials
- Acid spill- sodium bicarbonate
- Base spill- monosodium phosphate
- Small broom and dustpan
Anytime a spill occurs there are certain procedures to follow to ensure the situation is controlled as much as possible. If a small spill is not handled properly, it can turn into a much more serious situation. Large spills can turn into major catastrophes if not handled properly.
In the case of a chemical spill:
- Notify the CSO or PI. Dependent on the hazardous nature of the chemical spilled, the CSO or Principal Lab Supervisor will decide if the situation warrants notifying the UHD President, Health & Safety Officer, or any other responders.
- If the chemical is hazardous do not attempt to clean.
- Alert other staff and students in the area of the spill.
- Wear protective equipment, including safety goggles, gloves, and long-sleeved lab coat.
- Avoid breathing vapor of spilled materials.
- Absorb or cover the spill with suitable materials, collect residue, place in container and dispose of through the Campus Hazardous Waste Program (work with NS office and EH&S).
- Clean spill area with soap and water for final decontamination
- In case of skin contact, flush the area with large amounts of water for at least 15 minutes and seek medical assistance.
- In case of eye contact, flush eyes for at least 15 minutes with large amounts of water and seek medical assistance
- Remember to call 911 (x9-911) for all medical emergencies and notify the CSO and Principal Lab Supervisor. Dependant on the nature of the contamination, they in turn may need to notify the UHD President and Health & Safety Officer.
In the case of large chemical spills:
- Attend to injured or contaminated persons and remove them from the area and alert people in and around the lab to evacuate.
- Call 911 (or x9-911) for emergency responders (as needed) and have knowledgeable laboratory staff available to consult with emergency responders.
- Turn off ignition and heat sources.
- Close doors to affected areas.
- Meet at a designated area away from the incident to ensure all persons are accounted for.
- Notify the CSO and Principal Lab Supervisor who shall in turn notify: the University President, V.P. for Academic Affairs, Health & Safety Officer and the Director of the Physical Plant.
All experiments involving recombinant DNA, biological toxins, and other biohazards must be approved by the UH-System Biological Safety Committee. Approval can be obtained by filling out the Memorandum of understanding and Agreement (MUA) form found at the following: http://www.research.uh.edu/Forms/Biosafety-Committee.aspx. UHD will have a standing membership on the UH-System Biological Safety Committee until such time as there is a need to establish an independent Biological Safety Committee at UHD. In summary, faculty working with biological hazards must complete both and MUA (and get it approved by UH) and a PHA which must be approved by all levels at UHD.
Research Labs and Teaching Labs at UHD are designated as Biosafety Level 1 (BSL-1). BSL-1 is suitable for work involving well-characterized agents not known to consistently cause disease in immunocompetent adult humans, and present minimal potential hazard to laboratory personnel and the environment. BSL-1 laboratories are not necessarily separated from the general traffic patterns in the building. Work is typically conducted on open bench tops using standard microbiological practices. Special containment equipment or facility design is not required, but may be used as determined by appropriate risk assessment. Laboratory personnel must have specific training in the procedures conducted in the laboratory and must be supervised by a scientist with training in microbiology or a related science.
BSL-1 facilities are restricted to those persons whose presence is required for experimental or support purposes. Persons who are at increased risk of acquiring infection or for whom infection may be unusually hazardous are not allowed in BSL-1 laboratories. All equipment necessary for conducting experiments including centrifuges, incubators, water baths, etc., shall be kept in the BSL-1 restricted access laboratory. The PI establishes policies and procedures by which only persons advised of the potential hazards and who meet specific entry requirements (general health and training) may enter the laboratory. The PI has the final responsibility for assessing each circumstance and deciding who may enter or work in the laboratory.
Lower Containment Level Projects
Lower containment level projects may be carried out at the same time as BSL-1 projects. However, all personnel in the area must follow BSL-1 practices and procedures.
A universal biohazard sign with a BSL-1 designation is posted on the outside of all appropriate laboratories. The sign identifies the biohazardous agent(s), lists the name and telephone number of the PI and CSO, and indicates the special requirements for entry into and exit out of the laboratory.
The biosafety section of this manual is designed to serve as an overview of safety for students, staff and faculty. Personnel that are advised of special hazards are required to read and to follow specialized instructions on those practices or procedures.
Laboratory personnel must receive appropriate training on the potential biohazards associated with the work. Personnel must receive annual updates, or additional training as necessary for procedural or policy changes. Training for all personnel in the area will cover these requirements and any Standard Operating Procedures. The PI is responsible for insuring that, before working with organisms, all personnel demonstrate proficiency in standard microbiological practices and techniques and in the practices and operations specific to the laboratory facility.
The following are general guidelines for working under BSL-1 conditions.
- Always minimize entry/exit to/from the room. Plan your experiments so that once you enter the room, you stay in the room. Bring media, cells, protocols, notebooks, etc. with you. All manipulations involving biohazardous materials should be conducted in biological safety cabinets (biological hoods) or other physical containment devices within the laboratory.
- Minimize the generation of aerosols in experimental procedures.
- Load all centrifuge bottles, carriers, and tubes in the biosafety cabinet. Wipe outer surfaces with disinfectant before transporting.
- Remove and replace gloves often.
- Do not use contaminated gloves to handle common equipment.
- Plastic-ware should be substituted for glassware whenever possible. Extreme care must be taken with any contaminated sharp items, including needles and syringes, slides, pipettes, pipette tips, capillary tubes, and scalpels. Needles and syringes or other sharp instruments should be restricted in the laboratory for use only when there is no alternative and must be carefully placed in conveniently located puncture-resistant containers used for sharps disposal. Labs will contact that NS stock room when the containers are 3/4 full.
- Broken glassware must not be handled directly by hand, but removed by mechanical means such as a brush and dustpan available in the individual lab or in the NS Stock Room.
- Cultures, tissues, or specimens of body fluids are placed in two containers to prevent leakage during collection, handling, processing, storage within UHD.
- For shipping outside of UHD using a common carrier (i.e., Federal Express, UPS, DHL, etc.), the PI is required to attain any necessary permits and comply with all Department of Transportation / International Air Transportation Association regulations.
- All sterilization of contaminated materials is done in an autoclave. Biohazardous waste must be autoclaved weekly. Autoclaves must be tested periodically for proper temperature and pressure control. If you find that an autoclave is not operating properly, please notify the PI immediately
- All research materials must be decontaminated properly. All liquid or solid wastes must be decontaminated by autoclaving. Reusable materials such as glass media bottles should be autoclaved after decontamination with disinfectant.
- Tissue Culture dishes, plates, conical tubes and centrifuge tubes should be placed in autoclave bags. Also, disposable pipettes, pipette tips, contaminated gloves, Kimwipes, should be placed in the autoclave bag. All autoclave bags containing contaminated materials should be transported to the autoclave in a leak-proof container. Once the solid waste is autoclaved, it should be lowered into another trash bag and placed in the agreed area for pick up.
- Mechanical pipetting devices are to be used at all times.
- As is the case of all labs there should be no eating, drinking, smoking, handling of contact lenses, or applying cosmetics while in the lab. Food must be stored outside the work area in cabinets or refrigerators designated and used for this purpose only.
- Work surfaces shall be decontaminated daily and IMMEDIATELY following spills of organisms. All spills must be cleaned up immediately. Disinfectant soap is available for spills on skin. At the end of every working day, all work surfaces including equipment used (e.g., centrifuges and the Biological Safety Cabinet) are thoroughly wiped down with the appropriate disinfectant.
- Chairs used in laboratory work must be covered with a non-porous material that can be easily cleaned and decontaminated with appropriate disinfectant.
- Spills of biological materials are decontaminated, contained and cleaned up by appropriate professional staff, or others properly trained and equipped to work with concentrated infectious or potentially infectious material. Spills and accidents that result in overt exposures to biohazardous materials are immediately reported to the PI and the NS office. Medical evaluation, surveillance, and treatment are provided as appropriate and written records are kept. Any needle stick needs to be reported to the PI or NS office. For more details see section below on Procedures for Handling a Spill of a Biohazard.
- Biosafety Cabinet - Properly maintained Biosafety Cabinets (class II or III) are used for some manipulation of biohazardous materials (e.g., pipetting, dilutions, transfer operations, plating, framing, grinding, blending, drying, sonicating, shaking, centrifuging) except where equipment design provides for containment of the potential aerosol.
- Centrifuges - Use pressure seal tubes/bottles in centrifuge tube carriers. Wipe down the outer surface of the rotor with decontaminant before taking to centrifuge. If there has been any possibility of leakage, the inner walls of the centrifuge chamber and the rotor should be immediately decontaminated.
- Incubators - Use a dedicated biohazard incubator with tight fitting plugs to seal openings. The incubators in the facility are shared among several research groups, so be sure to label your containers. Clearly label any containers that contain infectious materials.
- Freezers/Refrigerators – Use a dedicated storage area for biohazards. There shall be a sign with the universal biohazard symbol on the freezer/refrigerator and materials shall be enclosed in a clearly labeled, unbreakable secondary container.
- Labeling - All equipment where biohazards are used or stored must be labeled with a universal biohazard symbol and the name of the agents used or stored.
- BSL-1 areas require special notations with respect to PPE.
- Lab Coats - Laboratory clothing that protects street clothing (dedicated cloth lab coat) must be worn in the laboratory and may not be worn outside the laboratory for any reason. Do not store any personal items, including overcoats, hats, etc. in the BSL-1 area.
- Gloves – Gloves worn while working with bacterial cultures, tissue culture cells and/or vectors. If a spill or splatter occurs, the hand will be protected after the contaminated glove is removed. Gloves are disposed of into the autoclave bag when contaminated, and replaced frequently during procedures. Do not wear contaminated gloves outside the work area. Keep common areas clean. Disposable gloves are not washed or reused.
- Face Protection - Goggles, mask, face shield or other splatter guards must be used for anticipated splashes or sprays of biohazards to the face.
Start-up Procedure for the Biological Safety Cabinet (Biological Hood)
- If your cabinet is equipped with a U.V. light, turn it off, and turn on the fluorescent light.
- Wipe down the work surface, interior walls (not including the supply diffuser), and the interior surface of the window with appropriate disinfectant.
- After placing your equipment inside the cabinet, close the sash to the proper operating height (normally eight inches but on some cabinets, ten inches). Check the owners’ manual for proper height. Avoid working in and operating the cabinet with the sash in any other position than the manufacturers recommended sash height.
- Wait another two to three minutes before working to clear all the contaminants from the work area.
- Use proper personal protective equipment as a barrier to possible exposures and to reduce contamination of your research. Wear a lab coat, gloves, and eye or face protection, if appropriate.
Working in the Biological Safety Cabinet
- The operator should be seated with the bottom of the sash level with his/her armpits.
- Perform all work using a limited number of slow movements, as quick movements disrupt the air barrier. Try to minimize entering and exiting your arms from the cabinet, but if you need to, do it slowly and straight in/out.
- To avoid excessive movements in and out of the cabinet, discard pipettes into a tray, container, or biohazard bag within the cabinet.
- Keep all materials at least four inches inside the sash opening.
- Plastic backed absorbent toweling can be placed on the work surface (but not on the front or rear grill openings). This toweling facilitates cleanup and reduces splatter and aerosol formation during a spill.
- All materials should be placed as far back in the cabinet as practical, toward the rear edge of the work surface and away from the front grille of the cabinet. Similarly, aerosol-generating equipment (e.g., vortex mixers, tabletop centrifuges) should be placed toward the rear of the cabinet.
- The general workflow should be from clean to contaminated (dirty). Materials and supplies should be placed in such a way as to limit the movement of dirty items over clean ones. Activities that create eddy currents (opening and closing doors, personnel walking near the cabinet), should be minimized as these types of activities can disrupt the air barrier.
- Open flames in a biosafety cabinet create a fire hazard and can damage the HEPA filter. If it is absolutely necessary to use a flame, a touch-plate micro burner equipped with a pilot light or a micro-incinerator may be used. Place the burner at the rear of the work area where the air turbulence from the flame will have the least possible effect on the air stream. The burner must be turned off when the work is completed.
- Aspirator bottles or suction flasks should be connected to an overflow collection flask containing appropriate disinfectant, and to an in-line HEPA or equivalent filter.
Completing the Work in the Biological Safety Cabinet
- All equipment that has come in contact with a biological agent should be decontaminated. The cabinet should be allowed to run for at least three minutes with no activity so that the airborne contaminants will be purged from the work area. Disinfect the surface of the equipment prior to removing it from the cabinet.
- After all items have been removed, wipe down all surfaces of the cabinet with a disinfectant, including the sides and back, and the interior of the glass.
Biological Safety Cabinet Certification
The cabinet must be certified upon installation, annually thereafter, and anytime it is moved (even if just across the room).
- Do not store equipment or supplies in the cabinet.
- Do not use the top of the cabinet for storage. The HEPA filter could be damaged and the airflow disrupted.
- Do not place items on the front or rear perforated grills. This reduces the airflow.
- Make sure the cabinet is level. If the cabinet is not, the airflow can be affected.
- Never disengage the alarm, as it indicates improper airflow, thereby effecting performance and endangering the researcher or the experiment.
At this time only cell cultures approved for work in BSL-1 laboratories may be utilized at UHD. The CSO will require PIs to provide verification that the cell lines they are currently working with can be handled in BSL-1 facilities.
Storage and retrieval of frozen cell cultures from liquid nitrogen requires appropriate personal protection equipment. The three major risks associated with liquid nitrogen (-196° C) are: frostbite, asphyxiation, and exposure. Gloves thick enough to act as insulation but flexible enough to allow manipulation of ampoules should be worn. When liquid nitrogen boils off during routine use of the freezer, regular ventilation is sufficient to remove excess nitrogen, but when nitrogen is being dispensed, or a lot of material is being inserted in the freezer, extra ventilation will be necessary.
There are currently two types of cultures that are utilized at UHD which could be classified as a biohazard. The culture types and the procedures for disinfecting them can be seen in Table 1 (below).
Type of Disinfectant
Means of Disposal
Eukaryotic cell cultures
Bleach (sodium hypochlorite)
Bleach before autoclave
Autoclave or dispose in the Biohazard waste
Unsued bacterial cultures
Bleach before autoclave
Autoclave or dispose in the Biohazard waste
Table 1. List of all potentially biohazardous materials utilized at UHD as of 2012.
- If a spill of biohazardous agents (cultures) occurs on a person in the lab, he/she should remove the contaminated articles or wash the exposed area with soap and water for at least 1 minute.
- If eye exposure occurs the eye wash station should be used per instructions. Medical attention should be sought as appropriate.
- If there is an injury or a near miss of an injury then a Report of Safety/Health Hazard and Near Miss must completed in the NS office.
Spills Inside the Biosafety Cabinet
- Move the glass shield down and wait at least 5 minutes to allow the Biosafety Cabinet to filter and clear aerosols.
- Wearing appropriate personal protective equipment at a minimum lab coat, safety glasses and gloves apply disinfectant for a minimum of 20 minute contact time directly on the spill and on all the potentially exposed surfaces of the cabinet.
- Wipe up spill with disinfectant soaked towels or other appropriate absorbent material. Wipe the walls, work surfaces, inside of sash and any potentially contaminated equipment with disinfectant soaked towels before removing it from the Biosafety Cabinet.
- Discard contaminated disposable material using appropriate biohazardous waste disposal procedures. Wipe down contaminated reusable items with disinfectant then place in autoclave bag or autoclave pans with lids for autoclaving.
- Those items that are non-autoclavable should be wiped down with disinfectant and kept wet for a minimum of 20 minutes before removal from Biosafety Cabinet.
- Remove protective clothing, when done and place in biohazard bag for disposal or autoclaving for reusable items.
- Run the Biosafety Cabinet for 10 minutes after clean-up before reusing.
- WASH HANDS!
Surface Spill in the Lab Outside the Biosafety Cabinet
- Be sure you are wearing appropriate personal protective equipment for size and nature of the spill to include at a minimum: lab coat, gloves and safety glasses.
- Place dry paper towels to establish a physical barrier between the spill and yourself. Then layer a second set of disinfectant-soaked towels over the spill.
- Starting from the outside and working in, carefully soak the spill with disinfectant being careful to minimize aerosolization.
- Decontaminate all items within the spill area. Wait at least 20 minutes disinfectant contact time to allow for adequate inactivation.
- Wipe equipment and reusable items with the disinfectant.
- Wipe up spill and discard contaminated disposables in the biohazard waste container.
- If sharps are present, use a mechanical device such as a dust pan and brush to pick up the sharps and place in an approved sharps container.
Spill Inside a Centrifuge
Wipe rotors and buckets with 70% ethanol, 1% SDS or 1% bleach.
Thoroughly disinfect inside of centrifuge with a minimum contact time of 20 minutes using a disinfectant that follows manufacturer’s recommendations and is effective for spilled agent. Dispose of contaminated materials in the biohazard waste container.
Spill Outside the Lab (in Transit)
Should a spill occur in a public area, do not attempt to clean up without the appropriate PPE.
Secure the area around the spill. Contact the NS Office, the CSO or your PI.
The release of a large volume of organisms (as a general rule anything over 10 ml or more) or a spill of material containing a high concentration of organisms must be reported to the PI or the CSO immediately. The PI and the CSO will with assess the risk and initiate an emergency response with the EH&S office as requested/appropriate. In the case of exposure a Report of Safety/Health Hazard & Near Miss form must be filled out. For students a Student/Visitor Accident Report Form will also have to be filled out by the PI or Chief Safety Officer.
Some laboratory courses at UHD are field based courses. Some research projects are also field based. In these cases it is important that field safety guidelines are followed.
Considerations Prior to Leaving
- Learn about potentially hazardous plants, animals, terrain, and weather conditions in the areas where field work will take place. This information can usually be obtained from sources such as state/national park websites, local collaborators, local residents, etc.
- Assemble safety provisions such as first aid kit and first aid manual (see below for recommendations), medications, allergy treatments, vehicle emergency kits.
- Try to recruit helpers. Using the “buddy” system in field work will greatly increase your safety.
- Ask your health insurance provider about coverage in the area you will be working.
Assemble a First Aid Kit and Medical Information
- A first aid kit should be taken on any off-campus field trip.
- A list of nearby hospitals should be obtained and taken into the field.
- The first aid kit should include the following: bandages, gauze, tape, antibiotic ointment, antiseptic swabs, mosquito repellant, medications such as acetaminophen, aspirin, antihistamines, and sunscreen.
- Every faculty and student planning to travel to the field must have completed all necessary travel arrangements in the NS office. All approved drivers should have a valid driver’s license and insurance information should be provided in vehicles.
- All vehicles should be equipped with working seat belts and mirrors and any other safety equipment/features required by law.
A variety of pests might be encountered in field work. Below are guidelines to minimize encounters with harmful pests.
- Keep garbage in rodent-proof containers and stored away from your campsite or work area.
- Shake all clothing and bedding before use.
- Do not set up camp near animal nests/dens.
- Look for pests before touching areas where they might live (wood piles, crevices).
- Avoid contact with sick or dead animals unless properly trained on how to handle them.
- Clothes with a tight weave should be worn and pant legs should be tucked into boots
- Use insect repellant.
- If bitten by a poisonous animal, seek medical treatment immediately.
- Educate yourself on local pests and poisonous animals/plants before leaving.
There are several species of poisonous or harmful plants and animals in Texas. Below is a summary of many of the common species likely to be encountered near Houston. If traveling outside of Houston or Texas, you should research other species you are likely to encounter at your destination.
- Spiders: black widows and brown recluses are poisonous. They can be found in shady, protected areas such as rock piles, under logs, outhouses and old buildings. The bite of these species will cause sweating, nausea, muscle cramps, fever and chills.
- Scorpions: these animals are usually active at night. Many are known to venture into campsites to feed on bugs attracted by light. They can be found in lumber piles, firewood, or old tree stumps. The sting is painful and can even be deadly.
- Bees and wasps: some species can be attracted by scents or food. They can inflict stings that can cause serious allergic reactions in some people.
- Ticks: the bites of ticks can spread diseases. To avoid ticks, wear clothing that covers as much skin as possible. Avoid brushing up against vegetation when possible. Always check yourself for ticks at the end of the day.
- Centipedes: these can be found under boards, in cracks and crevices and moist locations. They typically emerge at night. They can inflict a painful bite but it is not typically harmful.
- Snakes: there are several species of venomous snakes in Texas. To avoid most snakes, walk in open areas and do not put your hands under boards or in crevices where you cannot visually determine that there are no snakes present. If bitten by a poisonous snake, seek medical attention immediately.
- Poison oak and poison ivy: learn to identify these plants. They are very common problems in Texas and many people have an allergy to them. To prevent exposure, wear long pants and long sleeves. If exposure is suspected, wash the area with cold water and soap.
Exposure to certain pathogens is possible during field work. Below is a list of possible diseases that can be encountered and how to prevent them.
- Tetanus: puncture wounds, lacerations or burns can create a pathway for exposure to tetanus. Field workers should be sure that their tetanus boosters are up to date and should treat wounds to prevent tetanus.
- Rabies: several species are known to transmit rabies in Texas. Bites from infected animals can transmit this disease to humans. To prevent exposure, avoid contact with wild animals, particularly sick or dead ones. If your field work involves risk of being bitten by animals, you should consider getting immunized against rabies before the field work begins. If bitten by an animal, seek medical attention immediately, even if you have been immunized.
All students involved in research at UHD will be required to have safety training every semester they are involved in research. Students will be required to enroll in a 0-unit Safety and Compliance course through the NS Office (NS 3099). This will be true whether the student is doing research for units, as part of a paid program or even if they are volunteering. As part of the Safety and Compliance course students will be required to complete safety training. Faculty and Staff will be required to complete training specific to their area in accordance with state and federal guidelines. The PI is responsible for ensuring all staff and students are properly trained in the use of hazardous chemicals and equipment in the laboratory that is not covered in basic safety training. In these cases the PI must coordinate with the CSO about Laboratory training must include:
- Detection methods and observations that may be used to detect the presence or release of hazardous chemicals. Examples of detection methods include visual appearance, odor and an understanding of chemical monitoring devices.
- Physical and health hazards of the chemical.
- Work practices, personal protective equipment, and emergency procedures to be used to ensure that the employee may protect him/herself from overexposure to hazardous chemicals.
All laboratory workers must also be informed of the location and availability of the following:
- Location and information on how to read Material Safety Data Sheets (MSDSs).
- Reference material on chemical safety, including Material Safety Data Sheets (MSDSs).
- Permissible exposure limits (PELs) for OSHA regulated substances.
- Signs and symptoms associated with exposure to the hazardous chemicals found in the UHD labs.
The training of all staff and students shall be documented as to date, time, location, instructors name and what information was covered. Staff training records shall be maintained by the Health & Safety Office.
Medical consultation and/or examination by a licensed medical professional who is experienced in diagnosing and treating victims of chemical exposure, biohazard exposure or laboratory injuries will be made available to employees or students of UHD laboratories under the following circumstances:
- When any individual develops signs or symptoms associated with exposures to the hazardous materials being used.
- An accident such as a spill or equipment failure results in possible overexposure to hazardous materials.
- An accident such as a spill or equipment failure results in possible overexposure to hazardous materials.
- There has been exposure to a biohazard.
- There has been an injury in the field.
This would also include any follow-up exams deemed necessary by the licensed physician. All exams and consultations will be provided at no cost to the employee, without loss of pay, and at a reasonable time and place. UHD will provide the physician with:
- Identity of the hazardous chemical the employee was exposed to (if known).
- Description of the conditions under which the exposure or injury occurred.
- Description of the signs of exposure or symptoms of injury that the individual is experiencing.
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Last updated or reviewed on 5/9/13