Immediately clean after a spill or breakage. Hazards and safety measures associated with these gases are summarized in Sections 3.8 and 3.9. IATA: International Air Transport Association. µm) that floats on air currents for extended periods and can subsequently reach the pulmonary alveoli and small-droplet particles (>5 Dry ice can cause severe frostbite within seconds of direct contact. Use only face shields rated for short-wave UV light for this purpose. Pike RM. Paggiaro PL, Loi AM, Toma A. Implosion guards made of plastic mesh or plastic boxes have been used with suction flasks to contain glass pieces if the vessel fails. In 2008, CDC convened a Blue Ribbon Panel of laboratory representatives from a variety of agencies, laboratory organizations, and facilities to review laboratory biosafety in diagnostic laboratories. Perform daily cleaning of fluidic systems and sampler following manufacturer instructions. N Engl J Med 1996;335:1494–6. Past and present hazards of working with infectious agents. Keep instrument covers and panels closed and secure while instrument is in use. In general, veterinary diagnostic laboratories use biosafety level (BSL)-2 practices and facilities for general veterinary diagnostic work and do practical risk assessment of incoming accessions to determine whether decreased (BSL-1) or increased (BSL-3) biosafety practices or facilities are warranted. Simian immunodeficiency virus needlestick accident in a laboratory worker. Tube breakage during centrifugation presents the greatest risk for contamination because large aerosol clouds are produced. For routine work in veterinary diagnostic and clinical laboratories personal protective equipment (PPE) must be provided, used, and maintained in the laboratory workspace. Summary of packing requirements for exempt human specimens, Category B substances, and Category A substances, Leakproof primary and secondary containers, Pressure-resistant primary or secondary container, Absorbent between primary and secondary containers**, List of contents between secondary container and outer package, Quantity limits for either passenger or cargo aircraft, Shipper's Declaration for Dangerous Goods, Cost of labor and materials to pack substance. Could cause overpressure/rupture of a closed system. Do not hold in the hands while inoculating. http://nar.oxfordjournals.org/content/37/18/e119.full. Ensure instrument safety shields and containment devices are in place at time of use. 13.6.1. The inability to identify a specific event was also reported in a more recent study (27), which found that the probable sources of LAIs were apparent in only 50% of cases. In the hematology laboratory, the major causes of injuries are likely to be exposure to blood and body fluids; needle sticks, aerosols from centrifuge or removal of tube stoppers, tube breakage; or contaminated gloves (22). Rapid testing (direct molecular test kits). Biosafety and biosecurity in the veterinary microbiology laboratory and animal facilities. Surgical pathology risks are associated with manipulating large amounts of fresh tissues from unknown infectious sources, which may result in puncture, cuts, and splashes of blood and body fluids, similar to the autopsy where fresh organs must be viewed at a grossing table and cut into sections thin enough for fixation (i.e., "breadloaved"). The same concepts of demarcation and separation of molecular testing areas that are described in this section can be used to establish clean and dirty areas in other parts of the diagnostic laboratory. Heat flux into the cryogen is unavoidable regardless of insulation quality. Store at room temperature. The eyes are extremely sensitive to freezing, and liquid nitrogen or liquid nitrogen vapors can cause eye damage. Wayne, PA: Clinical and Laboratory Standards Institute; 2011. Human rabies prevention—United States, 2008: Recommendations of the Advisory Committee on Immunization Practices. The likelihood that an exposure will result in an LAI is directly related to several variables, including. Critical point dryers can be quite dangerous because of the high pressures generated within the "bomb" (. Clean any spills of body fluids or tissues immediately. J Natl Med Assoc 2004;96:1646–7. The most common agents of laboratory-acquired fungal infections are the dimorphic fungi Blastomyces, Histoplasma, and Coccidioides (18,19); most reported infections were caused by inhalation of conidia. Demonstrations of common problems or exposures that laboratory scientists might encounter in their normal work duties can be presented for individuals or groups. In non-microbiology sections of the diagnostic laboratory, the primary mistake may be assuming that a given specimen contains no infectious agents and then working with little attention to risk for infection. Clinical flow cytometric analysis of neoplastic hematolymphoid cells; approved guideline—second edition (CLSI document H43-A2). Determination of the extent of exposure, 14.3.3. The absence of hissing does not mean the vial is safe. The physical environment and space are appropriate for the work being done. The goal of a safety program is to lower the risk to as close as possible to zero, although zero risk is as yet unattainable as long as patient specimens and live organisms are manipulated. Such specimens received for testing need to be promptly and properly packaged and shipped to an appropriate reference laboratory. Install a sink equipped with either an automated motion-detecting faucet or knee or foot controls. Occupational Safety and Health Administration. The hazard to pathologists from tuberculosis specimens. Background to the 17 amendments adopted in the 13th revision of the United Nations Model Regulations guiding the transport of infectious substances. Lyophilized cultures are best maintained at 4°C or lower. Discard tissue remaining from culture activities of BSL-3 infectious agents into medical waste, and autoclave it. The high-velocity airflow can also aerosolize dried or liquid materials that might be present on the outside of the tube. Quality system essentials (QSE), definition, and related biosafety considerations. I. Yersinia pseudotuberculosis, • Fungi: Aspergillus fumigatus, Wear laboratory coat and eye protection when handling DMSO solutions. If ingested, dry ice can cause severe internal injury. Collins C, Grange J. Workers who handle or manipulate human or animal cells and tissues are at risk for possible exposure to potentially infectious latent and adventitious agents that may be present in those cells and tissues. Kyes K. Blood safety. Biohazards associated with cell culture reading, 8.3.4. Even after heat-fixing, the slide may contain viable tubercle bacilli and should be treated as contaminated. This is normal. Use special cryogenic liquid tongs when retrieving items from liquid nitrogen. Long-term preservation methods permit intervals of months or even years between subcultures. Two classifications of risk groups have been developed to facilitate the assessment of risk from various microbes and to recommend appropriate safety practices for the handling of those microbes (1). Xylene is categorized as a hazardous waste under RCRA and has been assigned EPA hazardous waste no. Rustigian R, Johnston T, Reihart H. Infection of monkey kidney tissue cultures with virus-like agents. NIH = National Institutes of Health; BSL = biosafety level; BSC = biological safety cabinet; PPE = personal protective equipment. Geneva: World Health Organization; 2004. Wear standard autopsy PPE. Arch Neurol 2009;39:103–7. Make sure that sharps containers are available in the work area, and inspect them periodically to ensure that they are never more than two-thirds full. Available at, Code of Federal Regulations. Manual removal of sealed caps; specimen aliquotting and pipetting. 13.2.1. Perform all work in a BSL-3 laboratory and within the BSC. When using a dedicated vacuum pump, many laboratories also include a suction flask containing coarse Drierite (W.A. The chemical hazards unique to the molecular virology laboratory include chloroform, ethidium bromide (Section 8.2.8) and guanidinium-based extraction reagents (Section 8.2.10). Use hand saws whenever possible to reduce aerosols. Wilkins D WA, Cossar YE, Miller DC. Healthcare personnel vaccination recommendations. Place the requisition in an outside pocket where it will avoid contamination with the specimen. Blastomyces dermatitidis (tissues), Coccidioides immitis (tissues), Wearing a respirator, such as N95, is highly recommended for protecting the laboratorian when processing and manipulating specimens or TB cultures. Available at, American Biological Safety Organization. Manipulation of infectious tissue can result in both airborne particles in a size (<5 Annu Rev Microbiol 1979;33:41–66. Batch culture systems for the mass production of micro-algae in 150 l cylinders. Routinely clean environmental surfaces before setting up the patient room/drawing station work area, and again before leaving the patient room or after each patient in the drawing station work area. The use of mercury in chemical analysis can be phased out in most, if not all parasitology laboratories. Clin Microbiol Rev 1997;597–610. Dispose of the container and unused contents in accordance with federal, state and local requirements. Document training and assess competency in use of and maintaining clean areas. Biosafety in Microbiological and Biomedical Laboratories (BMBL-5) and in the World Health Organization's Biosafety compendium on regulations and information sources. Additional precautions may be necessary if warranted by site-specific risk assessments. The more that laboratorians become aware of and adhere to recommended, science-based safety precautions, the lower the risk. Standard operating procedures should include. Patel R, Paya CV. For convenience, electrical outlets are recommended at each work station, along with telephone and computer jacks. Consult MSDS documents for more information. Government Printing Office (GPO), Washington, DC 20402-9371; Atlanta, GA: US Department of Health and Human Services, CDC. Only trained technicians are to service this equipment. Submission of fecal specimens in stool preservatives (commercially available vials) would decrease risks. Latent virus infections in primate tissues with special reference to simian viruses. Appropriate actions taken after an exposure can greatly reduce or even eliminate the chance that an exposure will result in an LAI. Do not use vinyl exam gloves for handling acetones and other aggressive organic solvents because vinyl gloves can be dissolved by these agents. Middleton PG, Miller S, Ross JA, Steel CM, Guy K. Insertion of SMRV-H viral DNA at the c-myc gene locus of a BL cell line and presence in established cell lines. † Some of these compounds can be found in common household, automotive, and industrial products, such as window, drain, toilet bowl, and surface cleaners, degreasers, antifreeze, and water treatment or swimming pool chemicals. Offices (e.g., of supervisors and laboratory director) that open into the clinical laboratory represent hybrid areas within the laboratory. Autoclave or disinfect all material coming into contact with cell cultures before discarding. Laboratory acquired infections. CDC. Toro C, Rodes B, Poveda E, Soriano V. Rapid development of subacute myelopathy in three organ transplant recipients after transmission of human T-cell lymphotropic virus type I from a single donor. Scanlon EF, Hawkins RA, Fox WW, Smith WS. These chemicals are strong irritants, and eye exposure can result in redness, irritation and pain. If frequent possible CJD cases are handled, laboratories may wish to dedicate an old microtome to this purpose. OIE risk groups and CDC/NIH biosafety level (BSL) classifications, Unlikely to cause disease; not considered infectious, Moderate individual and low community risk; unlikely to cause serious disease or be transmitted; effective treatment and prevention available, High individual and community risk; causes serious infections but not readily transmitted; effective treatment and prevention usually available, High individual and community risk; readily transmitted and no effective treatment or prevention available, Well-characterized agents not known to consistently cause disease in healthy adult humans; minimal potential hazard to laboratory personnel and the environment, Agents of moderate potential hazard to personnel and the environment, Indigenous and exotic agents that cause serious or potentially lethal disease as a result of exposure by the inhalation route, Dangerous and exotic agents that pose a high individual risk of aerosol-transmitted laboratory infections and life-threatening disease, Abbreviations: Recommendations for all laboratories include use of Class IIA2 biological safety cabinets that are inspected annually; frequent hand washing; use of appropriate disinfectants, including 1:10 dilutions of household bleach; dependence on risk assessments for many activities; development of written safety protocols that address the risks of chemicals in the laboratory; the need for negative airflow into the laboratory; areas of the laboratory in which use of gloves is optional or is recommended; and the national need for a central site for surveillance and nonpunitive reporting of laboratory incidents/exposures, injuries, and infections. Application of sodium hypochlorite solutions to spent solutions of EtBr will deactivate the ethidium bromide, but the reaction products are mutagenic, according to the Ames test (. Abbreviations: These activities can present a freezing hazard. Use acetones in a well-ventilated area (or chemical fume hood) to prevent respiratory irritation. US Department of Transportation, Pipeline and Hazardous Materials Safety Administration. The National Research Council, Committee on Hazardous Biological Substances in the Laboratory. Although the risks associated with antibiotic preparation and use are relatively low in the virology laboratory, antibiotic preparation and handling has been associated with hypersensitivity reactions and contact dermatitus (115,116) and asthma (116–120) in hospital, pharmaceutical, and animal workers. State and local disposal regulations may differ from federal disposal regulations. Manual of clinical microbiology, Eighth edition. (. Employers are to have a formal plan to address actions to be taken after an employee is exposed. MMWR 2004;53:281–4. Monthly safety mini-presentations can be presented during regularly scheduled laboratory meetings. Workbook for designing, implementing, and evaluating a sharps injury prevention program. Federal Register 2004;69:76044–76187 (. Release of oxygen gas, generally does not occur violently. Evans blue is used as a counterstain during fluorescence microscopy. A substantial charge can build up on the cap, and the charge must be relieved before touching the cap. Class 6 substances must be categorized into either Division 6.1 (toxic substances) or Division 6.2 (infectious substances). US Postal Service. Employee training can be accomplished by any of several methods, and nearly all of these can be adapted or combined to fit the needs of employees in a particular laboratory. For example: All proper behavior for avoiding or dealing with safety breaches such as those highlighted in these demonstration events should be described in a standard safety practices and procedures section in the laboratory safety manual. Large animal necropsy generally requires multiple prosectors working together in a way that will avoid accidental lacerations. Never put dry ice in beverages to cool them. Several manufacturers market safety devices to help remove caps from tubes and to break open ampoules (e.g., Current Technologies Saf De-Cap [Fisher Health Care, Houston, TX] and the Pluggo [LPG Consulting, Inc., Wood River, IL]). 2011. Sterilize a syringe by filling it, then emptying it with boiling water 3-4 times. 3.14.2. Darlow HM. Tissue that will be used for slide examination is fixed in formalin; subsequently, small tissue sections of interest are dissected with scalpels, placed in cassettes, replaced in specimen vats with formalin, and transported to histology. Consider all blood culture bottles as coming from patients potentially infected (e.g., with human immunodeficiency virus [HIV] or hepatitis), and handle appropriately. DMSO is a powerful solvent and can penetrate skin and latex gloves. Remove any broken glass in a spill area, and discard without contact with the hands (use [e.g.] Hopefully you will get the answer from text books or other online resources. The level of PPE use in routine veterinary diagnostic laboratory work should be sufficient for the BSL practices appropriate to the suspected or identified risk, and, at a minimum, include gloves and protective clothing. Documentation of biosafety training and continuous education is maintained in their personnel file and linked to their annual review. Many of these chemicals are dissolved in flammable solvents, and they must be kept away from heat and ignition sources. Mengling WL. All functions of the human and animal diagnostic laboratory — microbiology, chemistry, hematology, and pathology with autopsy and necropsy guidance — are addressed. Always use instruments according to manufacturer instructions. Use shrink seals or tape to seal petri plates (especially if mould begins to grow) in order to prevent accidental opening and spread of hyphal segments, conidia, or spores. Minimize or eliminate the use of multi-outlet power strips. Acetone is the principal organic solvent used in the virology laboratory and is primarily used as a fixative for cell smears. Because laboratory tests might be performed outside a traditional laboratory setting (e.g., doctor's office, outpatient clinic, community setting), these recommendations for training and education must be adapted to suit the employees performing the tests and the person who is overseeing them. For laboratories that contain multiple classes of BSCs, the hazards that are permitted to be manipulated within the specific unit need to be clearly indicated (by label) to staff (. Wear gloves, long-sleeved laboratory coat, and full-face shield when working with UV view boxes lacking protective filter shields. Guanidinium compounds are reactive with acids and other oxidizers, producing toxic fumes including cyanide vapors (thiocyanate and isothiocyanate derivatives), hydrochloric acid vapors (guanidinium hydrochloride), and nitrogen oxides (all forms). Use of disposable liners may reduce cleaning intervals of the equipment but does not replace the need to clean surface areas or equipment. To reduce exposures to The decision to use respirators in the laboratory may come either from the inherent risk of potential infectious agents in a clinical specimen (e.g., BSL-3 or risk group 3 agents) or from laboratory manipulations necessary for agent identification that may produce infectious aerosols. Allow the dry ice to sublimate or evaporate to the atmosphere in a well-ventilated area where CO vapor cannot build up. Franklin Lakes, NJ: Becton Dickinson; 2011. Laboratory exposures occur more often than is generally suspected. Transplantation 1994;58:852–4. Laboratory-associated infections: incidence, fatalities, causes, and prevention. If PCR testing fails to identify the virus, move the culture into the BSL-3 laboratory for further testing. State and local disposal regulation may differ from federal disposal regulations. Allow only one person to cut at a given time. Protecting People.™, of these potential hazards can be minimized by adoption of safe handling pra. Remove visible gross tissue from nondisposable instruments (knife handles, pans, aprons) before chemical disinfection or autoclaving. The three basic types of BSCs are designated as Class I, Class II, and Class III. provided as a service to MMWR readers and do not constitute or imply Care should be taken to collect all scraps of paraffin and unused sections on a disposable sheet. Cover the autopsy table with a plastic sheet to retain the majority of fluids, or alternatively, use tables with drains so that fluids may be collected in buckets or floor drains. Wipe off visible contamination by using a towel or gauze pad moistened with acceptable decontaminant, such as a 1:10 dilution of household bleach, or use the established laboratory disinfectant. Sterilize a quart (0.9 liter) jar, lid, and ladle. Shippers are responsible for being aware of these changes, adhering to current regulations, obtaining permits in advance of shipping, and interpreting applicable regulations for themselves and their facilities. J Neurol Sci 2000;177:154–6. Ensure that appropriate procedures for chain of custody are in place and followed even if the sample is not processed. Use of trade names and commercial sources is for identification only and does not imply endorsement by the U.S. Department of See Section 3.9 for additional information. PPE includes a variety of items, such as gloves, laboratory coats, gowns, shoe covers, boots, respirators, face shields, safety glasses, and goggles, that are designed to protect the laboratory worker from exposure to physical, biological, and chemical hazards. Zohar D, Luria G. A multilevel model of safety climate: Cross-level relationships between organization and group-level climates. Initiate a root cause analysis to determine all facts regarding the exposure, the root causes, and an action plan to correct identified causes. Use of scalpels, saws, and needles as well as exposure to sharp objects within the body, bone fragments, fractured metal, and/or needles, can result in cuts and percutaneous injuries. These containers are unacceptable for culture because the contents may have become contaminated. If you want me to write about any posts that you found confusing/difficult, please mention in the comments below. Wipe the liquid culture jar lid with Alcohol Wipes. Endogenous agents in primary cell cultures with special reference to latent viruses. Never place flammable liquids in a household refrigerator. Procedures such as centrifugation and cytospin processing can produce aerosols. Conduct an annual review of engineering controls because changes in practices and technology may put the laboratory staff at new risks. The low quantities used in some commercial reagents are relatively safe, but thimerosal exposure can have cumulative effects. Blood should always be handled carefully; avoid open cuts. Source: Adapted from OIE manual of diagnostic tests and vaccines for terrestrial animals, 6th edition. Dip cap or cork into molten paraffin wax to seal. Addendum III. Latex gloves will eventually dissolve in acetone, and double-gloving is recommended for short-term acetone usage. Allow appropriate contact time, then drain, rinse and dry. Clin Microbiol Rev 2001;14:659–88. Schaumburg, IL: American Veterinary Medical Association; 2008. The Occupational Safety and Health Administration (OSHA) defines PPE as "appropriate" if it does not permit blood or other potentially infectious materials to pass through or reach the employee's street clothes, undergarments, skin, eyes, mouth, or other mucous membranes under normal conditions of use (33). Part II: Immunization of adults. Use sturdy racks, buckets, or boxes that will ensure the item will remain upright.
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