US Environmental Protection Agency Office of Pesticide Programs Office of Pesticide Programs Microbiology Laboratory Environmental Science Center, Ft. Meade, MD Standard Operating Procedure for Handling Spills of Biohazardous Materials SOP Number: MB-13-05 Date Revised: 09-01-17 ------- SOP No. MB-13-05 Date Revised 09/01/17 Page 1 of 6 SOP Number MB-13-05 Title Handling Spills of Biohazardous Materials Scope The protocol presents guidelines for decontamination and cleanup of biohazardous spills. Application This SOP distinguishes between large spills vs. small spills, and spills inside vs. outside of the biological safety cabinet. Procedures for responding to a spill may vary, depending upon the degree and location of the spill of biohazardous material. Approval Date SOP Developer: Print Name: SOP Reviewer Print Name: Quality Assurance Unit Print Name: Branch Chief Print Name: Date SOP issued: Controlled copy number: Date SOP withdrawn: ------- SOP No. MB-13-05 Date Revised 09/01/17 Page 2 of 6 TABLE OF CONTENTS Contents Page Number 1. DEFINITIONS 3 2. HEALTH AND SAFETY 3 3. PERSONNEL QUALIFICATIONS AND TRAINING 3 4. INSTRUMENT CALIBRATION 3 5. SAMPLE HANDLING AND STORAGE 3 6. QUALITY CONTROL 3 7. INTERFERENCES 3 8. NON-CONFORMING DATA 3 9. DATA MANAGEMENT 3 10. CAUTIONS 4 11. SPECIAL APPARATUS AND MATERIALS 4 12. PROCEDURE AND ANALYSIS 5 13. DATA ANALYSIS/CALCULATIONS 6 14. FORMS AND DATA SHEETS 6 15. REFERENCES 6 ------- SOP No. MB-13-05 Date Revised 09/01/17 Page 3 of 6 1. Definitions 1. Appropriate disinfectant = EPA-registered hospital disinfectant with a label claim for the class of microorganisms (e.g., vegetative bacteria, spore formers, viruses, fungi, mycobacteria) being disinfected. All disinfectants must be used according to the directions (e.g., use dilution, contact time, etc.) specified on the label. 2. Spill = A spill is defined as a biohazardous material out of control. The quantity of the biohazardous material spilled is not the sole determining factor in deciding whether or not an event is classified as a spill. Rather, the essential issue is whether the biological agent, the location, and the quantity collectively cause the situation to be beyond the control of the laboratory worker. A major spill is one that cannot be handled safely by laboratory employees in the immediate area. A minor spill is one which can be handled by the laboratory workers in the immediate area without posing a serious threat to their health and safety, and that can be cleaned up with available absorbents and disinfectants Additional abbreviations/definitions are provided in the text. 2. Health and Safety Follow procedures specified in SOP MB-01, Laboratory Biosafety. The Study Director and/or lead analyst should consult the Material Safety Data Sheet for specific hazards associated with any disinfectants. 3. Personnel Qualifications and Training Refer to SOP ADM-04, OPP Microbiology Laboratory Training. 4. Instrument Calibration Not applicable 5. Sample Handling and Storage Refer to SOP MB-22, Disinfectant Sample Preparation. 6. Quality Control Not applicable 7. Interferences Failure to become familiar with and to put into practice the procedures set forth in this SOP will result in analysts who are a danger to themselves, others, and the environment. 8. Non-conforming Data Strict adherence to the biosafety practices is required. Nonconformance will result in notification, retraining, or disciplinary action of laboratory employees. 9. Data Management The Branch Chief is responsible for documenting accidents and exposures ------- SOP No. MB-13-05 Date Revised 09/01/17 Page 4 of 6 associated with spills. 10. Cautions 1. Lack of use or understanding of this SOP may negatively impact the decontamination efforts of laboratory staff and hence cause unnecessary exposure of employees to human pathogenic microorganisms. 2. Failure to clean the ultraviolet lamps in the BSCs will reduce the lamps' effectiveness. Periodically clean the ultraviolet lamps in the biological safety cabinets (BSCs) with a lint-free cloth dampened with alcohol. 3. If a liquid sodium hypochlorite (bleach) solution is used to decontaminate stainless steel surfaces (e.g., BSC) following a spill, be sure to wash the surface with water, 70% ethanol, or an EPA-registered disinfectant to remove excess sodium hypochlorite. 11. Special Apparatus and Materials 1. Autoclave. 2. Trash bags (clear in color, autoclavable) or containers inside and outside of the biological safety cabinets for collection and storage of biohazardous waste. 3. Personal protective equipment (PPE) such as gloves, safety glasses, lab coats, disposable laboratory garments, shoe covers, and temporary clothing (i.e., scrubs). 4. Biosafety Spill Kit containing items such as gloves and tongs for handling broken glass, dustpan/brush, shoe covers, disposable lab coat, and safety glasses. 5. Signage to identify biohazardous materials and to limit access to laboratories. 6. Appropriate EPA-registered hospital disinfectant/tuberculocide. 7. Bleach solutions made fresh as needed. Discard solution at the end of the day. The original container of bleach will be discarded six months from the date of receipt or designated as use for cleaning only. a. Use 1:10 diluted bleach solution at neutral pH for decontamination purposes (spore-forming microorganisms). Using an EPA registered sodium hypochlorite product containing at least 6% sodium hypochlorite, dilute as follows: 1 part bleach, 8.4 parts water, and 0.6 parts 5% white vinegar or 5% lab grade acetic acid. 8. Key card readers are used to limit access to testing laboratories. Only ------- SOP No. MB-13-05 Date Revised 09/01/17 Page 5 of 6 authorized personnel are permitted to enter. 12. Procedure and Analysis 12.1 Guidance for Spills of Biohazardous Material - Reporting Instructions a. Accidents are handled according to the practices outlined in this subpart, as well as procedures referenced in the Occupant Emergency Plan (OEP) and the ESC Chemical Hygiene Plan (CHP). b. Report all spills and accidents, regardless of how minor a spill to the Branch Chief and the SHEM manager (or call security desk at extension 5-2800). c. The analyst is responsible for reporting (via email) spills to the Branch Chief. d. The Branch Chief and SHEM manager will determine if additional written documentation or follow-up is warranted. 12.2 Recommendations for Reducing Potential for Spills of Biohazardous Material a. Use secondary containment (e.g., autoclave bin) when transporting live cultures in liquid or solid media. b. Use secondary containment to store biohazardous waste that is generated during the course of an assay. c. Prepare the least amount of culture necessary for an assay. d. Maintain a clean, well-organized work environment. 12.3 Biohazardous Organisms Requiring Biosafety Level 1 and 2 Containment a. Refer to Attachment 1: Guidance for Spills of Biohazardous Organisms Requiring Biosafety Level 1 and 2 Containment - Spills Outside and Inside the BSC. 12.4 Decontamination of Cloth Lab Coats, Street Clothing, and Footwear a. Decontaminate clothing with an appropriate disinfectant or by autoclaving. b. If using disinfection as a means of decontamination, treat area of contamination and surrounding area with an EPA- approved disinfectant, following the label-specified dilution and contact time. c. Autoclave clothing potentially contaminated with microorganisms in spore form. Use a 3 hour liquid cycle as per SOP MB-01. ------- SOP No. MB-13-05 Date Revised 09/01/17 Page 6 of 6 d. It is less harmful to clothing to autoclave it in a tray than it is to bag it. Do not put water in the tray with the lab coat. Rather, put a second tray into the autoclave and add water to this tray. e. After clothing is decontaminated (by disinfection or autoclaving), immerse it in water containing detergent to aid physical removal of decontaminated biohazardous material. f. Rinse lab coat and dry then set aside to be sent out with the lab coat laundry service. g. Take street clothing and footwear home and launder. 13. Data Analysis/ Calculations None 14. Forms and Data Sheets 1. Attachment 1: Guidance for Spills of Biohazardous Organisms Requiring Biosafety Level 1 and 2 Containment - Spills Outside and Inside the BSC 15. References 1. Centers for Disease Control and Prevention and National Institutes of Health, 2009. Biosafety in Microbiological and Biomedical Laboratories, 5th edition. U.S. Department of Health and Human Services. U.S. Government Printing Office, Washington, D.C. ------- |