Contaminated laboratory waste Laboratory waste should be separated into colour-coded containers. The recommended colours are;Yellow - for incineration Light blue or transparent with blue inscription - for autoclaving (but may be incinerated subsequently Black - normal household waste: local authority refuse collection White or clear plastic- soiled linen There are three practical methods of treating contaminated laboratory waste: sterilization by autoclaving, chemical disinfection, and incineration. The first two are laboratory processes; incineration involves the transport of material off-site. Sterilization and disinfection are not synonymous: sterilization implies the killing of all microorganisms; disinfection kills most microorganisms but depend greatly on the chemical used. Autoclaves Autoclaving involves the timed exposure of materials to steam above atmospheric pressure and hence at temperatures above 100oC. Autoclaves operate at high pressures and temperatures and their manufacture, installation, and use are regulated. In the UK, they are subjected to the Pressure Systems and Transportable Gas Containers Regulations 1989, which requires regular inspection and maintenance and regular checks on the effectiveness of the sterilization cycles. For most purposes, sterilization is achieved by exposure of material to 121oC for 15-20 min. Higher temperatures such as 126oC for 10 minutes, or 134oC for 3 minutes are permitted. Temperatures are monitored by thermocouples in modern autoclaves. Chemical and biological indicators are also widely used, the most well-known of which is the "autoclave" tape. Biological indicators is common use consist of strips or discs of filter paper which have been soaked in suspensions of Bacillus stearothermophilus spores, then dried and packed in porous envelopes or sachets. These are then placed in various positions in the load and after processing, the strips and discs are removed from their packing and placed in nutrient broth which is incubated for 24-48 hours. The presence of turbidity indicates failure to sterilize. It was customary to place material for autoclaving In wire baskets and cylindrical buckets. While these containers are perfectly acceptable for bottled fluids, they are not suitable for potentially infected material since they do not contain spilled material. Therefore, shallow buckets or other

They are not compatible with cationic detergents. Laboratory uses include discard jars and disinfection of surfaces. wood. Most activators contain a dye so that the user can be sure that the disinfectant has been activated. Iodophors and quaternary ammonium compounds (QAC) are more popular in the US than in the UK. spores.the active ingredient is chlorine. Hypochlorites may cause irritation of skin. enveloped viruses (including Lassa and Marburg). Chemical Disinfection Chemical disinfection should be regarded as a first line defence especially in the case of discarded bench equipment. such as sodium bicarbonate. and both enveloped and non-enveloped viruses. Mycobacteria and non-enveloped viruses are less susceptible and spores are generally resistant. and lungs. There is a rough spectrum of susceptibility of microorganisms to disinfectants. They are not compatible with cationic detergents. or as a liquid. spores. and plastics. Clear Phenolics . fungi. paraformaldehyde.formaldehyde (gas) and glutaraldehyde (liquid) are good disinfectants. For spillages of blood and discard jars which may receive much protein. fungi. 2500 ppm is recommended. 1. They are active against vegetative bacteria (including mycobacteria). Both forms are heated to liberate the gas which is used for disinfecting enclosed spaces such as safety cabinets and rooms. Most phenolics are active in the presence of considerable amounts of protein but are inactivated to some extent by rubber. For general purposes and discard jars. The most susceptible are vegetative bacteria. Hypochlorites are considerably inactivated by protein and to some extent by natural non-protein material and plastics. fungi. Aldehydes have a more limited application and alcohol mixtures are less popular. The hypochlorites sold for laboratory applications in the UK contain 100000 ppm. Aldehydes ."solid-bottomed containers" should be used for sterilizing infected material which may leak. 10000 ppm should be used. formalin. Autoclavable plastic bags are now in general use and they must be permeable to steam.these compounds are effective against vegetative bacteria (including mycobacteria). eyes. They are inactive against spores and non-enveloped viruses. They should not be used on the metal parts of centrifuges. The most commonly used disinfectants in laboratory work are clear phenolics and hypochlorites. Hypochlorites decay rapidly in use and diluted solutions should be replaced after 24 hours. The mouths of plastic bags should not be closed before autoclaving since this will reduce steam penetration. and both enveloped and non-enveloped viruses. Clear phenolics should be used at the highest recommended concentration. and enveloped viruses. Formaldehyde is supplied either as a solid polymer. which should be followed by autoclaving or incineration. fungi. They are active in the presence of protein and are not very much inactivated by natural or man-materials or detergents. Dilutions should be prepared daily and diluted phenolics should not be stored for more than 24 hours. Hypochlorites . Effectiveness and stability after activation varies with product and the manufacturer¶s literature should be consulted. Glutaraldehyde usually requires an activator. . Laboratory uses include discard jars and surface disinfection but they corrode some metals and thus care is necessary. 3. Skin and eyes should be protected. It is important to ensure that the container used is not overlarge since it may lead to inadequate sterilization. 2. which is very effective against vegetative bacteria (including mycobacteria).

Full-face respirators should be worn when rooms are being fumigated with formaldehyde. Alcohol and alcohol mixtures . and respiratory tract. They may also be used in discard jars but they are comparatively expensive.they are cationic detergents that are effective against vegetative bacteria and enveloped viruses and some fungi. albeit slowly. Precautions in the use of disinfectants . Disposable gloves and safety spectacles. They are inactivated by protein and a variety of natural and plastic materials and soap. 4. Aldehydes are toxic and precautions must be taken when formaldehyde is used.discard jars should be robust and autoclavable. Quaternary ammonium compounds . Discard jars should be washed with hot water before reuse. They are rapidly inactivated by protein. and non-enveloped viruses. It is important that discard jars are filled frequently with known active dilutions of disinfectant. They are usually used for cleaning surfaces and are also very popular in food hygiene laboratories because of their detergent nature. They are not inactivated by protein and other material or detergents. these iodines are effective against vegetative bacteria (including mycobacteria). some natural and plastic substances and are not compatible with anionic detergents. They are relatively harmless to skin but may cause eye irritation. or a visor should be worn by anyone handling strong disinfectants. . They are left open on the bench but are closed at the end of the working session. Re-usable pipettes . spores. goggles. Wide-moth screw-capped polypropylene bottles or jars are better still. eyes. They can also be used for handwashing since they usually come with a detergent. Funnels may be used to prevent splashing and aerosol dispersal. Their laboratory uses are therefore limited but they have the distinct advantages of being stable and non-corrosive to metals. They should remain in disinfectant for at least 18 hours before washing and/or autoclaving. fungi. Discard Jars . Laboratory supervisors should ensure that inappropriate articles are not placed in discard jars e. 6. Iodophors are relatively harmless to the skin by some eye irritation may occur. Laboratory uses include discard jars and disinfecting surfaces. Iodophors . Polypropylene beakers or jars are probably the most serviceable items. The contents of the discard jars should then be autoclaved and/or incinerated. It is better still to autoclave the jar before chorine compounds. some disinfectants have undesirable effects on the skin. 5. spores. re-usable pipettes should be completely immersed in disinfectant so that no air remains. Glass jars should not be used because they are easily broken.alcohol and alcohol mixtures are effective. a 25ml pipette has no place in a small 1 litre jar.As indicated above.g.after use. They are non-toxic and harmless to the skin and eyes. They are not active against mycobacteria. against vegetative bacteria and enveloped viruses.Glutaraldehydes are particularly useful in disinfecting metal surfaces as they do not cause corrosion. and both enveloped and no-enveloped viruses. Alcohols and alcohol mixtures are useful for disinfecting surfaces. They are not effective against spores. not overloaded with protein or floating articles. fungi and non-enveloped viruses.

from mopping up to total decontamination of the room by fumigation 4. breakage or other accidents involving infectious material.Disinfection after accidents There is an immediate need for disinfection after spillage. which disinfectant to use. After a spillage.g.this is of particular concern when it happens in reception rooms with clerical staff. Laboratory workers should not bend down to inspect the damage.breakage of tubes in a centrifuge can disperse large amounts of aerosols. Leakages should be dealt with by a member of the laboratory staff. This should be left for at least 30 minutes before the towels are removed. the safety cabinet should be disinfected by formaldehyde. all accidents can be treated as above for decontaminating . the centrifuge buckets and rotor can be removed and autoclaved and the bowl disinfected. 3. The tray used should be flooded with disinfectant and left for several hours. Any possibly contaminated clothing should be removed. which may not be repeatable may be considered as important as protecting the staff. and the extent of the room decontamination procedures e. post office. After the room is considered safe to enter. If material is spilled in a safety cabinet. Simple spillages . Leakage into transport boxes should be treated in the same way except that the box can be autoclaved. centrifuge accidents . The action to be taken is much the same as that for the breakage of a culture. If that is not practical. all people in the room should hold their breath and leave. Instead. Appropriate equipment should be available for all such emergencies. Disposable gloves should be worn and the offending container and any others which may be contaminated be placed on a tray which is then placed in a large plastic bag and removed to a laboratory. public service vehicle or aircraft. preferably a safety cabinet. The safety officer should draw up protocols for decontamination procedures after particular accidents. Sealed buckets should be used for all Hazard Group 3 agents. Serious spillages . There is much splashing and a considerable amount of aerosol is dispersed.these may range from leakage of a pathological specimen in a hospital van to leakage or breakage of a culture in a public place. Action to be taken ranges from simple mopping up with disinfectant to decontamination of large items of equipment or of whole rooms. the fan should be left running while the spill is being dealt with. The surface on which the leakage occurred should then be covered with paper towels over which the appropriate disinfectant is poured. The towel should removed after 30 minutes and the are swabbed with fresh disinfectant. 5. Hypochlorites should not be used because it will bleach the fabric. Saving the specimen. In principle. Transport associated accidents . then the affected area should be sponged with a disinfectant-detergent. The safety officer should by this time have ascertained the possible nature of the organism involved and decide what respiratory or other protection is required when re-entering the room. The door should be closed and a warning sign posted onto it. leakage of specimens . The area should then be swabbed with fresh disinfectant and left for a further 30 minutes before it is dried. 1. 2.the most serious spillages are those that involve cultures dropping on the floor and break.simple spills should be covered promptly with a paper towel and disinfectant poured gently on the towel. Thirty minutes is a reasonable time to allow the aerosol to settle or clear.

surfaces and rooms. The instrument is then flushed through with a disinfectant recommended by the manufacturer. He should leave the room immediately after starting the reaction. The operator should wear eye and face protection and if formalin is used. After decontamination. It is safe to pour most specimens down a sluice or deep sink which will join the public sewer. the effluent discharge tube should be taken at least 25 cm into he laboratory waste plumbing system or that it is trapped in a bottle. Many companies require certificates that state specifically that the equipment is microbiologically safe. wet disinfection and fumigation. ventilators. The interiors of incubators. 2. It may also be possible to disinfect the whole room with a sprayer. Decontamination Procedures 1. Wet disinfection is only applicable if the floors do not allow liquids to leak into the rooms below and any remaining furniture or equipment is not likely to be damaged by disinfectant. Protocols for dealing with these emergencies have been worked out by most health departments and have been published by the WHO. In the case of automated equipment. then it is of utmost importance to ensure that there is no possible leakage from the room. Disposal of contaminated liquid waste Small amounts of contaminated liquid waste may be poured into disinfectant but problems arise with large amounts such as urines. It should be washed out several times with clean water and dried. it is essential to inform the senior medical microbiologist and environmental health officer at once. Fumigation of whole rooms is now not generally recommended because of health threats. before a change of use or before building alterations or may be necessary to disinfect a room after a spillage of infectious material. Two methods may be used. pipe runs and cracks should be sealed with masking tape. the equipment should be taped or otherwise secured so that it cannot be used and labelled. 3. effluents from automated equipment.laboratory protective clothing should be autoclaved before it is sent for laundering or placed in "hot wash" bags which should not be opened in laundries until they have been exposed to very hot water. refrigerators and similar cupboard-like equipment should be swabbed thoroughly with glutaraldehyde or alcohol-formalin mixture and left overnight. The door should then be locked and sealed with masking tape and a warning notice be posted and the room left for 24 hours. breathing apparatus should be worn. However. Rooms . The floor is mopped by a disinfectant and then left wet for at least 30 minutes. The operator must have full eye and face protection as well as a respirator. Windows. If fumigation is carried out. Laboratory protective clothing . Equipment . and sink and lavatory effluents from Maximum Containment is desirable to have a regular procedure for decontaminating equipment that is to be serviced on site or returned to a manufacturer for any reason. since the potential for the spread of infection is so great. Sewage disposal plants deal quite .

The safe disposal of the various effluents from Containment Level 4 laboratories is a design consideration. The effluents from automatic apparatus are unlikely to be very hazardous since microorganisms are likely to have encountered inimical chemicals on their way through the machine. are scrubbed with an approved enzymatic cleaner and thoroughly rinsed. then tested for the presence of pathogens before discharge into the sewers. neat disinfectant (hypochlorite or phenolic) should be added to the urine. The only hazard to the operator is from splashing and aerosol production when pouring and this should be done with care. there is often confusion about how to properly maintain and use such equipment. milking. inspection. wrapping. H. If it is known or suspected that the urine contain Hazard Group3 pathogens. What follows is a brief discussion of the essential steps for ensuring patient safety by properly sterilizing instruments. packaging. Afterwards or at the end of the day. the sluice or sink should be flushed with disinfectant. Established ambulatory surgery centers generally have both central sterile supply areas and staff members trained to properly clean and sterilize instruments. there can be confusion about what must be sterilized and a lack of knowledge about the proper steps required to appropriately clean and sterilize instruments. Precautions Against Hepatitis and HIV Proper sterilization ore and more minor surgeries are being performed in ambulatory care settings. Pre-cleaning needs to be done at the point of use to prevent the drying of organic material. They must be sure all instrument surfaces. critical items-any object that enters sterile tissue or the vascular system-must be sterilized. A competency tool can be found here. cleaning. Even where tabletop sterilizers are readily available. Per E. This includes most surgical instruments used in the ambulatory setting. Staff should wear appropriate PPE and follow approved procedures to prevent BBP exposure. These include pre-cleaning. This can be accomplished by either wiping the instrument with a wet cloth or placing the instrument in an enzymatic cleaner. Many steps are required to ensure that instruments are appropriately sterilized. chemical treatment is preferred. In other ambulatory care settings. autoclaving and maintaining the autoclave.effectively with most pathogens. however. If an ultrasonic . including hinges. Sometimes. Spaulding's classification scheme. and that means there is a greater need for staff in these areas to reprocess instruments. Cleaning includes many steps. The effluents are usually taken to one or one holding tanks where they are heated by steam for a predetermined time.

instruments with moveable parts should be immersed in a milk solution and removed without rinsing. A steam indicator should be placed in the center of the pack with one end visible when the pack is opened.cfm?Section=Infection_Connection&Template=/CM/Conten tDisplay. chips or worn spots. Staff should be able to verbalize the recall/resterilization procedure in case of failure of the biological indicator. All instruments should be in the open position with any curved tips pointed in the same direction. It is important to ensure that hinged instruments open easily and that the jaws are properly ensuring that packs are loaded in a manner that allows for free steam and air circulation. steam indicators that do not appropriately change color. All instruments should be inspected for cracks. A competency checklist can be found at http://www. instruments should be wrapped in a single pouch of an appropriate size. A competency checklist can facilitate teaching and ensure compliance with these standards. After inspection. Autoclaving should be done following manufacturer's directions. and package integrity concerns or compromised storage and handling conditions. The packet should be secured with steam indicator tape and labeled with the date of sterilization. Any instruments found with defects should be removed from service and sent for repair. A log should be kept that details each time the autoclave is run. All instruments should be inspected before wrapping. Sharp instruments should be inspected for sharpness. we have seen real improvement with the cleaning and sterilization process .cfm&ContentFileID=2103. Milking: After cleaning. Since implementation of this checklist.apic. care should be used to be sure all manufacturers' recommendations are followed. visible condensation seen in a package. the load number and the initials of the person preparing the package. every time a biological indicator is sent and every time maintenance is performed.machine is used.