It is the responsibility of the Principal Investigator to identify potential biohazards and to specify safe practices and procedures. All laboratory personnel must be informed of the potential hazards and trained in safe handling techniques. Service personnel and cleaning staff that enter the facility must be informed of the hazards or potential hazards that might be encountered. The following Biosafety Practices outline the minimum requirements for all research and teaching laboratories using biological commodities. Issues of non-compliance will be reported to the appropriate Chair, Dean and the Chair of the Institutional Biosafety Committee for their action. § GOOD MICROBIOLOGICAL PRACTICE The use of aseptic techniques and other good microbiological practices achieves three very important objectives. These are: OCCUPATIONAL HEALTH: The prevention of illness, disease or injury when working with these commodities. ENVIRONMENTAL SAFETY: The prevention of contamination of the laboratory by the organisms being handled. PRODUCT PROTECTION: The prevention of contamination of the work with organisms from the environment. The first is of prime importance as regards working safely whereas the second and third are key considerations in relation to the quality of the research. The principles of good microbiological practice should be applied to all types of work involving biological commodities (including genetic modification work) irrespective of containment level. In addition aseptic technique is also invaluable for preventing contamination of tissue cultures. Aseptic technique is based on creating a special (clean) micro-environment in which to grow and keep the micro-organism of interest and prevent all contact with the outside world. This micro-environment is usually some sort of culture or holding vessel such as a flask, bottle or petri-dish and the organisms can either be on a solid agar medium or be suspended in a broth, diluent or other fluid medium. The principles of aseptic technique are: Prior to use all components of the system (the inside of the vessel, the medium and any objects used in the manipulative processes) must be sterile; and In the inoculation, incubation and processing steps particular care must be taken to avoid cross-contamination. This involves: Keeping the vessel closed except for the minimum time required to introduce or remove materials; working with a bunsen burner and flaming the opening of the vessel (passing it quickly through the bunsen flame) whenever tops are removed. The upwards current of hot air created by the bunsen prevents contaminated air or particles dropping into the culture vessel when the lid is open. Using manipulation techniques that minimise any possibility of cross contamination eg: opening lids with the little finger so that tops are not put down on the benches; and Ensuring that all of the objects that may come into contact with the culture , such as loops and pipette tips, are sterile before use, are not contaminated by casual contact with the bench, fingers or the outside of the bottle etc during handling and are decontaminated or disposed of in a safe manner immediately after use.   § STANDARD MICROBIOLOGICAL PRACTICES The following requirements are required for any laboratory using infectious or toxic agents: Prohibit mouth pipetting. Use devices that are accurate and safe. Eating, drinking, smoking, storing food or utensils, applying cosmetics and inserting or removing contact lenses are not permitted in any laboratory. Long hair must be tied back or restrained. The laboratory is kept neat, clean, and free of materials not pertinent to the work. Work surfaces must be cleaned and decontaminated with a suitable disinfectant at the end of the day and after any spill of potentially dangerous material. Intensive cleaning must be done at regular intervals. Members of the staff should wash their hands after handling biological commodities known or suspected to be contaminated, even when gloves have been worn, animals, and when leaving the laboratory. All techniques and procedures are performed in a way that minimizes the creation of aerosols and splashes. . BSCs are used whenever there is a potential for aerosol/splash creation or when high concentrations/large volumes of infectious agents are used. Decontaminate all contaminated liquid or solid materials before disposal or reuse; Transport contaminated materials safely; properly covered, and labeled, that need autoclaving or incinerating at a site away from the laboratory. Contaminated materials that are to be autoclaved should be transported to the autoclave in durable leakproof containers (e.g. autoclave bins), which are labeled as biohazardous. Wear laboratory coats, gowns or uniforms only in the laboratory. Do not wear laboratory clothing in non-laboratory areas. Biosafety Program recommends solid front laboratory coats (wrap arounds) and color when working inside of laboratory and white if protecting contaminated inner clothes when transferring to another laboratory. Disinfect contaminated clothing by appropriate means. Suitable footwear with closed toes, non-absorbable and preferably with non-slip soles must be worn in all laboratory areas. Bare legs are not acceptable when working with infectious materials. Safety goggles, face shields, face masks, or other personnel protective equipment are made available when necessary to protect the eyes, face, and body from splashes, impacting objects, harmful substances, UV light, or other rays. Safety face and eye wear (goggles) must be worn when necessary to protect the face and eyes from It is also recommended that eye protection be worn when operating a microwave oven or flame looping. Only persons who have been advised of the potential hazards and meet any specific entry requirements are allowed to enter the laboratory working area; laboratory doors must be kept closed when work is in progress; access to animal houses are restricted to authorized persons; children are not permitted into laboratory working area. There is an insect and rodent (vector) control program. Animals not involved in the work being performed are not permitted into the laboratory. The use of hypodermic needles and syringes must be restricted to parenteral injection and aspiration of fluids from laboratory animals and diaphragm vaccine bottle. Avoid autoinoculation and the generation of aerosols during use and disposal. Needles should not be recapped, bent, cut, sheared, but should be promptly placed in a puncture-resistant container and sterilized before disposal. Wear gloves for all procedures that may involve direct or accidental contact with blood, infectious materials, or infected animals. Gloves removed aseptically and autoclaved with other laboratory wastes before disposal. Gloves should be appropriate size, type, etc., for the individual. Gloves should be decontaminated with other laboratory wastes before disposal. Report all spills, accidents, and overt or potential exposures of infectious materials immediately to the laboratory supervisor. Prepare a written accident report. Maintain a written record and submit these to the Biological Safety Officer. Provide appropriate medical evaluation, surveillance, and treatment. Collect and store baseline serum samples from all laboratory and other at-risk personnel. Periodically collect additional serum specimens depending on the agents handled or the function of the facility. The laboratory supervisor must provide laboratory safety training. Adopt working safety or standard operation procedure's manual that identifies known and potential hazards and specifics practice, procedures, and operations that minimize or eliminate such risks. Advise personnel of special hazards and require personnel to read and follow this internal laboratory standard operating practices and procedures. Keep laboratory windows and doors closed when active manipulation is in progress. This controls the spread of aerosols throughout the building and avoids accidental exposure to unannounced visitors. Annually certify required equipment such as; Biological Safety Cabinets, Laminar Flow Clean Benches, autoclaves, Pressure vessels, etc., please call the Biological Safety Program for more details. Have biological indicators attached to all biohazards wastes being autoclaved to determine sufficient sterility. Log all autoclave runs. Shipping of biological agents or materials must conform with the latest U.S. Federal Standards and private courier regulations.   § STANDARD OPERATING PROCEDURES These standard operating procedures are a minimum set of guidelines for the handling of biological commodities. Individual administrative units, laboratories, or research groups are expected to develop more detailed procedures as their situations warrant. Supervisors and principal investigators are responsible for complying with and/or enforcing appropriate safety and hygiene measures in the work areas they supervise. Some rules or standard operating procedures, which apply to all work areas on campus, include the following: General Procedures Respect and understand the safety and health hazards associated with the biological commodities and equipment you use, and practice the following general safety guidelines at ALL times: Accident response: If an injury requiring emergency medical assistance has occurred, call 6-911. Report to your pre-designated medical advisor. Biological spills: If a toxic/hazardous chemical has made contact with the skin, start flushing the area immediately. If emergency assistance is required, call 6-911. Children and unauthorized persons: Children and other unauthorized persons should not be in laboratories where hazardous materials or hazardous equipment are being used. Disposal of chemicals: see Lab Safety Program Electrical: Access to electrical equipment (e.g. plugs, switches and electrical panels) should be maintained free from obstructions to allow immediate access in an emergency. All receptacle outlets in laboratory spaces should be the polarized grounding type. Ground Fault Circuit Interrupters (GFCI's) should be used in those locations involving wet processes or outdoor work, including electrical outlets within six feet of sinks. All electrical hand tools used inside laboratories should be grounded or double insulated. All electrical extension cords used should be visible and inspected on a periodic basis for damage and/or defects. Cords should not run in aisles or corridors where they might be damaged or create a tripping hazard. Cords should not be run through doors, walls or partitions, under rugs, or above dropped ceilings. They should not be wrapped around fixtures, tied in knots, or draped over pipes, lights, or ventilation ductwork. Extension cords should not be used as substitution for fixed receptacle outlets. Cords used for 110-120 volt service should be UL listed standard heavy-duty three-wire equipped with a polarized three prong plug. Two-wire type extension cords should not be used. Emergency eye wash/safety showers: Be certain safety showers/emergency eyewashes are properly located and maintained. These units should be located in areas that will be immediately accessible (reachable within 10 seconds). There should be no obstructions that might inhibit the use of this equipment. Eye washes and safety showers should be flushed on a regular basis to verify that the units are working and to clear the lines of stale water and debris. Whenever these emergency units are checked for proper functioning, written documentation showing the date and person's initials performing the check, should be maintained. Equipment: Use proper equipment that is in good condition. For example, never use chipped or cracked glassware. Shield pressurized or vacuum apparatus and safeguard against bumping or overheating. Fire extinguishers: Fire extinguishers must be available, charged, and hung in a location that is immediately accessible (reachable within 10 seconds). There should be no obstructions that might inhibit the use of this equipment. Make sure that all extinguishers are checked annually. Each extinguisher should have a tag indicating the date it was last checked. Food, drink, and cosmetics: Eating, drinking and the application of cosmetics are forbidden in areas where hazardous chemicals are used and should be done only in well-defined, designated non-chemical areas. Do not store food in the same refrigerator with chemicals, biohazards or radioactive materials. Horseplay: Practical jokes or other behavior that might confuse, startle, or distract, another worker is forbidden when hazardous chemicals are present. Housekeeping: Exits, aisles and safety equipment must NOT be obstructed in any way with equipment, furniture, or other items. Aisles within the laboratory should be 36 inches in clear width. Work areas and floors are not to be used for excessive storage. Doors which are not in use but which are accessible from a corridor or adjacent room should be appropriately labeled if they are blocked on the interior of the room. Hallways are not to be used as storage areas. Mouth pipetting: Mouth pipetting is forbidden. Signs: Laboratories where biohazardous materials or operations are present must post a sign with the universal biohazard symbol. An emergency contact card, updated at least annually, should be posted on each laboratory entrance. Smoking: No smoking in laboratories. If you have been using biological commodities, be sure to wash your hands before smoking. Unattended experiments: If operations involving hazardous biological commoditiies are carried out with no one present, it is the responsibility of the worker to design procedures to prevent the release of hazardous biological commodities in the event of interruptions in utility services such as electricity, cooling water, and inert gas. Lights should be left on, and signs should be posted identifying the nature of the operation and the hazardous substances in use. If appropriate, arrangements should be made for other workers to periodically inspect the operation. Similarly, if unattended experiments require the use of running water, the worker should develop procedures to make sure the experiment is checked periodically for water leaking from the system. Working alone: When working with hazardous materials, it is advisable to have a second person present, or at a minimum, maintain contact via telephone.   § PERSONAL PROTECTIVE EQUIPMENT (PPE) and PERSONAL HYGIENE Personal protective equipment (PPE) and personal hygiene are basic aspects of laboratory safety. Wearing appropriate personal protective equipment and practicing good personal hygiene as described below will minimize exposures to hazardous chemicals during routine use and in the event of an accident. Attire: Wear a lab coat or apron, cover legs (no shorts or skirts) (Biosafety prefers the use of solid-front laboratory coats) and feet (no sandals, open-toed shoes or absorbable materials), confine loose clothing and long hair. Nylons and/or pantyhose are not recommended because they may melt upon contact with acid and absorbable. Eye protection: Personnel including students, staff and visitors in laboratories wear safety glasses, goggles, or face shields at all times where eye hazards are a possibility. Goggles are recommended when impact hazards are possible, i.e. use of microbial loops. Contact lenses may be worn in the laboratory; however, they do not provide any protection of the eyes. Persons who wear contacts must use the same eye protective equipment as persons who do not wear contacts. Face shields: Full-face shields must be worn when conducting a procedure which may result in a violent reaction. Full-face shields with bottom caps to protect the neck are preferred because they provide the best protection. Gloves: Gloves are essential when working with hazardous substances. The proper gloves will prevent skin absorption, infection or burns. Glove materials vary in effectiveness in protecting against chemical hazards. Leather or a similar type of protective gloves should be used when inserting glass tubing into stoppers, make sure to lubricate the tubing and wear the protective gloves to prevent being cut in the event of the tubing slipping and breaking. Personal hygiene: Hands should be washed frequently throughout the day, after glove removal, before leaving the lab, after contact with any hazardous material, and before eating, drinking, smoking, or applying cosmetics. Respiratory protection: Work in a biological safety cabinet or provide adequate ventilation when working with materials that produce aerosols or when using chemicals concurrently with biologics. If the use of a respirator is required, you must comply with EHSO policy, which includes a medical assessment, fit testing, and instructions on proper use.   § HAZARD ASSESSMENTS For each task involving hazardous materials or physical hazards, a written hazard assessment must be conducted and SOP written. Supervisors and Principal Investigators are responsible for conducting hazard assessments. Such assessments should not be limited to chemical hazards, but should also include such issues, when applicable, as radiation hazards, biological hazards, heat and cold hazards, and physical hazards. Evaluate the effects of possible exposure to a hazardous agent (e.g., symptoms of clinical infection, allergic symptoms or complaints related to pharmacological effects of end products, by-products, medium components or inactivated biological agents) detect changes in employee health that may indicate the need for a change in job procedures or job assignment detect patterns of disease in the work force or evaluate the effectiveness of control measures.