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					SERI #23 - SPECIAL ANIMAL SAFETY PROTOCOL FOR                                                    ROUTE OF
DIPHTHERIA TOXIN IN MICE                                                                         ON:

TOXIC – IN NON-IMMUNIZED HUMANS, MINIMAL LETHAL DOSE                                             IP INJECTION

Investigator Name:                                             Date of Administration(s)
IACUC #:                                                       Date(s):______ _______ __________
Emergency Phone #:
Date:                                                          Date(s): ______ _______          _________

Investigator              Research staff will inform animal care staff     Signature of research staff who notified
Responsibility-           ahead of time that diphtheria toxin will be      Animal Facility:
                          used, and arrangements will be made for
Notification of           housing of animals. Fresh cages will be used     _________________________
Animal Care:              for the animals at the time of administration.   Date of Notification:
                          The animals will be housed at BL-2 until the
                          first cage change.                               _________________________
Investigator’s  It is highly recommended that anyone working with the Diphtheria Toxin
Responsibility- be immunized.
Basic           • Toxin must NOT be worked with in a dried state. It should be pre-diluted in
Precautions         the bottle and sharps precautions must be used during injections. All excess
                    toxin must be returned to the lab and disposed of as hazardous chemical
                     waste. The powdered toxin must be carried down to the animal facility in its
                     original sealed container and labeled secondary containment.
                • Animal bedding is not to be changed for at least 2 days after the date of
                    administration of the Diphtheria Toxin.
                • Disposable gown, double gloves, surgical mask and safety glasses in
                    addition to the usual protective clothing must be worn.
Investigator’s  • This SASP will be posted on the door of the room in which the animals will be
Responsibility-     housed.
Posting         • Cages will be labeled with a RED CARD denoting diphtheria toxin along with
Requirements:       the date of administration. SPECIAL CARCASS label should be placed on
                    the permanent cage card and the specific agent being injected should be
                    written on the back of the cage card.
Investigator’s  Diphtheria Toxin will be administered by IP Injection on an absorbent pad on
Responsibility- the bench top. Luer lock syringes recommended. Follow dosage and
Administration precautions in Standard Operating Procedure (SOP). The first cage change
and Hazardous after each drug administration is to be done by the Research Staff no sooner
Waste Cage      than 2 days after the administration. The bedding is considered contaminated
Change:         and requires special handling.
                • Wearing two pairs of gloves and a surgical mask, transfer the animals to
                      clean cages.
                • Animals must remain in the BL2-N Room.
                If live animals are temporarily removed from the BL-2 room (no sooner
                than 2 days after administration):
                 • You must secure the cages in such a way that if they fall, their contents will
                      not spill.
06/30/10 FINAL, Revised 11/19/10
                           •    A rubber band, removable blue tape or bungee cord must be wrapped
                                around the cage. These items are stocked in the BL2 room.
                           • No implements used in the BL-2 room may leave the BL-2 room.
                           • You must fill out a new SASP and post it at the entrance of the temporary
                                work area where you are manipulating the BL2 animals.
                           • The temporary work area needs to be disinfected when the animal is
                                returned to the BL-2 room.
                           • Remove the SASP from the temporary work area door, secure cages with
                                rubber bands or tape and return animals to the BL2 housing area.
                           • All waste generated during the procedures, must be handled as regulated
                                medical waste. You must follow BL2 PPE practices.
                          If this is the end of diphtheria toxin administration remove RED CARD and
                          place in dirty cage. Printed side up.
                          NOTE: If The administrations will continue write diphtheria toxin on a
                          blank RED CARD and place face up in cage. The original RED CARD stays
                          on the cage.
                          • Replace the tops, food hoppers, water bottles on the soiled cages and place
                              in a heavyweight YELLOW plastic bag.
                          • Remove outer gloves and place in bag before sealing bag.
                          • Twist the ends of full bags, goose-neck, and tie off.
                          • If diphtheria toxin administration is complete: Remove the sign from
                              the door of the room housing the animals.
                          • Transport closed bag to designated rack and AF staff will process
                          After this first cage change there is no need for further special precautions to
                          be taken regarding the animals or the cages as long as the animals have not
                          received any more diphtheria toxin.

Investigator’s and        •    After the first cage change standard husbandry practices should be utilized
AF’s Responsibility            for animal handling.
AF’s Responsibility-      Animal care technicians transport the bags of soiled cages to the BSC HEPA
Waste                     filtered dumping station on the dirty side.
Disposal:                 • Colored bags are not opened until there is space to process the cage.
                          • Place new hazwaste bag in dump station for only this hazwaste disposal.
                          • Put on second pair of gloves along with standard PPE.
                          • Dump bedded cages into cage changing station. Dump food. Dump water.
                          • Wipe out cages with paper towel to remove all particulate debris.
                          • Discard the paper towels into the dump station.
                          • When all of diphtheria toxin cages and food hoppers are empty
                               immediately place into cage washer.
                          • Dispose of paper towels used for wiping in the bag. Close bag and bring to
                              hazardous waste boxing station.
                          • When all cages are finished, wipe down cage dump station with disinfectant
                              strength chlorine dioxide.

AF’s Responsibility-      Dispose of carcasses as regulated medical waste.

06/30/10 FINAL, Revised 11/19/10
06/30/10 FINAL, Revised 11/19/10

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