CCM Approval: IACUC Approval:

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1 CCM Approval: IACUC Approval: 1.0 Purpose: The purpose of this SOP is to describe the necessary procedures for the safe administration of potentially hazardous agents 1 to rodents via injection, drinking water, and/or food when housed at CCM-managed facilities. The goals are threefold: (1) To ensure the toxic agent is administered safely within the animal facility, (2) To ensure clear notice of the toxic agents in use for the benefit of those working in the vicinity of the cage, and (3) To ensure proper clean-up and disposal of all waste associated with the work. 2.0 Scope: The scope of this SOP applies to all CCM-managed facilites and all staff involved in associated inoculation and animal care procedures. Staff with responsibilities: 2.1 Facility supervisory staff 2.2 Animal care staff 2.3 Cage wash staff 2.4 Investigative staff 3.0 Definitions: N/A 4.0 Equipment/Supplies: 4.1 Agent-specific Safety Data Sheets (SDS) 4.2 Biosafety Cabinet or Chemical Fume Hood 4.3 Cardboard Biohazardous Waste/Incinerate Box (i.e. burn box) 4.4 Disinfectant in Spray Bottle (e.g., Quatricide T.B., Virkon S, Rescue) 4.5 Dump Station 4.6 Enclosed Transport Container 4.7 Hazardous/Toxic Agent card (See Figure 1, below.) 4.8 Low-profile Sharps Container 4.9 Personal Protective Equipment (PPE): Disposable gloves or gauntlets (double-glove with chemically resistant gloves) Face shield, goggles or safety glasses (for chemical-splash protection) Facility-Standard Protective clothing (e.g. isolation gown and sleeves) Face mask or N-95 mask (if fit-tested annually) 4.10 Spare Clean Rodent Cage(s) 1 Refer to BWH toxic agents Appendix 2016 BWH-APD-RES-Toxic Agents 2016 BWH-SOP-RES-502 V02 Page 1 of 6

2 5.0 Procedures: 5.1 Researchers must complete the following prior to working with chemical hazards in lab animals: Conduct an agent risk assessment by reviewing the SDS and pertinent data (see Appendix A for risk assessment form) Ensure agent is included in the approved BWH insight protocol under BWH hazardous agents form Notify the animal facility manager before starting and review procedures, location of supplies, and chemical hazard dirty cage return location. 5.2 Research staff will perform the following steps utilizing full-sterile technique: Preparation and transport of inocula Prepare the inocula in the research lab in a chemical fume hood or biosafety cabinet (BSC) with up-to-date certification. Ensure that any precautions identified in risk assessments are oberved while preparing agents for administration Transport the inocula to the animal facility in securely sealed tubes or syringes that are contained in an appropriately labeled, enclosed secondary container that is shatterproof and sanitizable (smooth and impervious) and has been packed with sufficient packing material to absorb the fluid in the event of a spill. Include the chemical name and the hazard on the label Deliver the container of inocula to the designated supply entry airlock of the animal facility and disinfect the container following the usual spraying-in procedures, being careful to not upend the container of inocula Unpacking the hazardous agent container Retrieve inocula from the designated supply entry airlock and bring it to a procedure room that is appropriately equipped for this work. Turn on the chemical fume hood or biosafety cabinet for at least five minutes before use Don two pairs of gloves and eye and/or face protection, if required by risk assessment requirements Spray the work surface of the chemical fume hood or the BSC with virucidal disinfectant (e.g. Virkon S, Rescue) If required, spray a low-profile sharps container with disinfectant and place in the hood or cabinet. BWH-SOP-RES-502 V02 Page 2 of 6

3 CAUTION: To ensure proper airflow in the cabinet, do not cover the downdraft grates Outside the cabinet, spray all surfaces of the transport container with a virucidal disinfectant Remove the sealed tubes or syringes from the transport container Remove the transport container from inside the cabinet. Note: Hood must be designated for chemical hazard use. The hood may not be shared until its surface is fully cleaned and the chemical hazard removed Administration of chemical Note: Follow the appropriate steps below depending on the route of administration Administration via water bottle: Wear the PPE described above Place the clean cage inside the hood or cabinet and remove the water bottle from the cage and remove the stopper from the bottle Add the agent to the bottle Place the stopper on the bottle Transfer subject rodents from their original cage to the clean cage using full sterile technique inside the hood or cabinet Return the treated chemical hazard bottle to the new cage Administration via chow: Note: Chow must be formulated at the point of manufacture Follow steps above as described, but place special diet in food hopper and leave water bottle untreated Administration via injection or gavage: Wear the PPE described above; place a clean, empty cage inside the hood or cabinet Place the cage containing the subject animals inside the hood or cabinet Serially remove each subject rodent from the cage, inoculate it, and then place into the clean cage. Use needle-safe or safety sharp devices to prevent personal injury Dispose of the needle and syringe in the sharps container. WARNING: Do not recap needles Cage labeling for hazard communication Affix the Hazardous/Toxic Agent card so it is the most outward facing cage card. BWH-SOP-RES-502 V02 Page 3 of 6

4 Record the name of the agent, route of administration, treatment date(s), and legible name and contact information of the person responsible for the animals Indicate end date if multiple treatments are planned or hazardous substance is administered in water Clean-up following administration of hazardous chemical Return the resealed tubes or syringes of inocula to the transport box. Unused inocula must return to the laboratory of origin for purposes of disposal or future use. Note: Storage and disposal of the agent must follow Environmental Affairs guidelines Spray the interior surfaces of the cabinet with disinfectant spray, wipe with disposable towel and discard into waste bin. Note: Chemical hazard spills must be handled according to Environmental affairs instructions. Hazard must be wiped from common area and materials used to wipe up hood disposed of by incineration (use biohazard plastic bags) Remove outer gloves and discard in the waste bin. 5.3 CCM animal care staff only change hazardous agent cages if they meet spot change criteria (i.e. excessively wet or soiled) as outlined in BWH-SOP-HUS-221, Performing Daily Rodent Checks. Follow procedures below in Section 5.4 for cage changing procedures. 5.4 Cage change and clean-up must be performed 72 hours after final hazardous agent administration by research staff Using BSC and facility PPE with double nitrile gloves, transfer rodents to clean cage Prepare dirty cage for return to cage wash chemical hazard return rack: Dump food from hopper into dirty cage bottom Pour all remaining water from the dirty cage bottle into the soiled bedding and on food for absorption Replace food hopper, filter top, and empty water bottle Keep chemical hazard card displayed on dirty cage Spray down the interior of the hood or cabinet and external dirty cage with disinfectant and wipe dry Remove outer pair of gloves and discard in trash Deliver LABELED dirty cages to the designated Chemical Hazard Cage Return in the facility for processing by CCM staff. 5.5 Dirty cage processing by CCM cage wash staff BWH-SOP-RES-502 V02 Page 4 of 6

5 Cage wash personnel must don the following PPE when processing chemical hazard cages in addition to their normal uniform: Nitrile gloves Goggles or face shield N95 mask Isolation gown Using facility dump station, empty chemical hazard cage contents into burn box Immediately send cage components through tunnel/rack/bottle washer for sanitation Deliver waste to designated area for pick-up by waste management for incineration. Figure 1 Hazardous/Toxic Agent 6.0 References: 6.1 BWH-SOP-HUS-221 Performing Daily Rodent Checks 6.2 BWH-SOP-HUS Microisolator Cage Changing Using Full Sterile Technique Document History Version Revisions Date 00 New Standard Operating Procedure 08/01/ Added Appendix A Laboratory Risk Assessment reviewed by 02/28/2017 safety 02 Significant revisions: chemical hazard cages dumped using engineering controls in cage wash 06/13/2017 BWH-SOP-RES-502 V02 Page 5 of 6

6 Appendix: A Laboratory Risk Assessment When hazardous agents are identified, a risk assessment should be performed by all staff handling the agent. The laboratory risk assessment (LRA) is one method to ensure agents are being used safely. At a minimum, staff working with hazardous agents should: 1. Identify the hazard 2. Assess the risk of hazard 3. Implement controls to mitigate the risk of hazard To identify the hazard, always review the Safety Data Sheet (SDS) of your material. In most cases, section 2 of the sheet will assist in evaluating what the hazards are. A rating of Category 5 is the lowest hazard, Category 1 is the greatest hazard. Once the hazard has been identified, assess how likely it is that you may be exposed. Ask questions such as: Is this chemical volatile? (will it get in the air) How am I preparing this chemical? What is the volume of the material being used? What are effects of an exposure to the material? How severe are the affects of an exposure, and what is the likelihood of exposure? What will be done if you are exposed to the material? Once assessing the risk of exposure, you should determine how to prevent an exposure. Examples of methods to prevent exposures are: Lab coats will reduce chance of exposure to skin or clothing Fume hood will remove vapors/fumes and provide barrier Biosafety cabinet will remove particles and provide barrier Gloves will provide a temporary barrier For very hazardous agents, they should be used in a designated area, away from others For hazardous agents, biosafety cabinets or chemical fume hoods should always be used. These devices will engineer the hazard away from the user. After you have implemented all controls, assess to determine if the process is safe and get help if you have doubts about personal safety. Additionally, discuss what a response would require in the event of an accidental exposure or chemical spill. Table For Completion: Material: Formaldehyde Hazard: Toxic Carcinogen Risk: Volatile, small volume 5ml Controls: Fume hood, gloves, lab coat, goggles Effective: Yes/No, if No, reevaluate controls or contact EH&S. Spill: Page 15000, Occupational Exposure: BWH-SOP-RES-502 V02 Page 6 of 6

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