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Animal Biosafety Level-2 (ABSL-2) Rodent Husbandry, Study notes of Biosafety

Sul Ross State University (SRSU)Biosafety

A class II biosafety cabinet. Page 2. SOP #408.15. ABSL-2 Rodent Husbandry. Effective 3/22. Page 2 of 6. (BSC) is utilized for cage change-outs and procedures, ...

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STANDARD OPERATING PROCEDURES
DIVISION OF COMPARATIVE MEDICINE
UNIVERSITY OF SOUTH FLORIDA
SOP#: 408.15 Date Issued: 10/98 Date Revised: 3/22 Page 1 of 6
TITLE: Animal Biosafety Level-2 (ABSL-2) Rodent Husbandry
SCOPE: All Authorized Personnel
RESPONSIBILITY: Facility Manager and Technical Staff
PURPOSE: To Outline the Proper Procedures for Safe Husbandry Practices
for Rodents Housed at ABSL-2.
I. PURPOSE
1. To outline the proper procedures for safely conducting husbandry of rodents housed
under animal biosafety level 2 (ABSL-2) conditions.
2. Reduce the risk of exposure of research and animal care staff to biohazardous
agents within the animal facility.
II. RESPONSIBILITY
1. The Facility Manager is responsible for ensuring:
a. That staff contributing to husbandry are adequately trained to perform the
husbandry practices described.
b. Implementation of the procedures described.
c. Dedicated ABSL-2 rooms for infectious agents, and animal housing rooms for
rDNA and PDX biocontainment at the cage level are clearly labeled with the
biohazardous agent present and specific safety practices implemented.
d. Appropriate personal protective equipment (PPE) is available.
e. Biohazard Safety Data Sheets (SDSs) are accessible, when available.
f. Safety practices have been communicated to the relevant personnel.
2. It is the responsibility of the animal care staff to:
a. Read, understand, and follow the procedures described.
b. Don appropriate PPE and review room signage and SDS prior to
implementing the procedures described.
III. PROCEDURES
1. Containment practices are in accordance with Institutional Biosafety Committee
(IBC) protocol-specific requirements for the agent in use.
2. Typically, containment of replication deficient viral recombinant DNA constructs or
primary human patient-derived xenografts (PDX) administered to mice or rats is at
the cage level in a conventional animal housing room with (A) signs SC #247 and
SC #251 posted on the outside of the door, (B) all primary enclosures administered
such agents identified with an orange biohazard cage card flag indicating agent
administered, exposure date, release date, PI, and emergency researcher contact
information, and (C) grouped together on the rack. A class II biosafety cabinet
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STANDARD OPERATING PROCEDURES

DIVISION OF COMPARATIVE MEDICINE UNIVERSITY OF SOUTH FLORIDA

SOP#: 408.15 Date Issued: 10/98 Date Revised: 3/22 Page 1 of 6

TITLE: Animal Biosafety Level-2 (ABSL-2) Rodent Husbandry SCOPE: All Authorized Personnel RESPONSIBILITY: Facility Manager and Technical Staff PURPOSE: To Outline the Proper Procedures for Safe Husbandry Practices for Rodents Housed at ABSL-2.

I. PURPOSE

  1. To outline the proper procedures for safely conducting husbandry of rodents housed under animal biosafety level 2 (ABSL-2) conditions.
  2. Reduce the risk of exposure of research and animal care staff to biohazardous agents within the animal facility.

II. RESPONSIBILITY

  1. The Facility Manager is responsible for ensuring: a. That staff contributing to husbandry are adequately trained to perform the husbandry practices described. b. Implementation of the procedures described. c. Dedicated ABSL-2 rooms for infectious agents, and animal housing rooms for rDNA and PDX biocontainment at the cage level are clearly labeled with the biohazardous agent present and specific safety practices implemented. d. Appropriate personal protective equipment (PPE) is available. e. Biohazard Safety Data Sheets (SDSs) are accessible, when available. f. Safety practices have been communicated to the relevant personnel.
  2. It is the responsibility of the animal care staff to: a. Read, understand, and follow the procedures described. b. Don appropriate PPE and review room signage and SDS prior to implementing the procedures described.

III. PROCEDURES

  1. Containment practices are in accordance with Institutional Biosafety Committee (IBC) protocol-specific requirements for the agent in use.
  2. Typically, containment of replication deficient viral recombinant DNA constructs or primary human patient-derived xenografts ( PDX ) administered to mice or rats is at the cage level in a conventional animal housing room with (A) signs SC #247 and SC #251 posted on the outside of the door, (B) all primary enclosures administered such agents identified with an orange biohazard cage card flag indicating agent administered, exposure date, release date, PI, and emergency researcher contact information, and (C) grouped together on the rack. A class II biosafety cabinet

ABSL-2 Rodent Husbandry Effective 3/ Page 2 of 6

(BSC) is utilized for cage change-outs and procedures, or if a changing station is utilized, additional PPE is required. Eye shielding and mask are required when working outside a BSC.

  1. In contrast, typical containment of replication competent infectious agents administered to mice or rats is accomplished in a separate, dedicated ABSL- containment room with (A) sign SC #252 on the door indicating the infectious agent(s) used within (B) primary enclosures are identified with an orange biohazard cage card flag and (C) grouped together on the rack. All cage change- outs and procedures involving infectious agents are performed in a BSC in the ABSL-2 animal housing room.
  2. Before entering the room, assemble all supplies necessary to complete task.
  3. Access to rooms housing ABSL-2 rodents is limited to essential personnel. Doors are kept closed when experimental animals are present, and when appropriate, locked when the room is unattended.
  4. Husbandry and care of ABSL-2 rodents are completed after other colonies.
  5. Before entering the room review ABSL-2 signage posted on the door for PPE requirements and specific practices implemented for the protocol/agent.
  6. When ABSL-2 rodents are not housed in a separate room (e.g., primary enclosures of mice administered recombinant DNA or PDX), ABSL-2 cages shall be grouped together and individually identified with an orange biohazard cage card flag indicating the agent in accordance with SOP 006.
  7. Don a disposable gown, gloves, sleeves and shoe covers prior to entering the room. Additional PPE as required by signage must also be donned, which may include but is not limited to eye protection, face shields, and respirators.
  8. Work surfaces are cleaned and decontaminated with Oxivir Tb) before and after use.
  9. Cage changing of infectious agent administered rodents must be performed within a BSC.
  10. Cage changing of rDNA or PDX administered rodents should be performed within a BSC (see SOP 1127 ) during the IBC interval of agent containment (e.g., first change after 3 or 7 days for replication deficient adeno- or lenti-viral recombinant DNA constructs, respectively) when present or whenever there is a high potential for creating aerosols. Eye shielding and mask are required when working outside a BSC (e.g., changing station). As a rule of thumb: a. Intervals of cage change-outs will be determined based on the associated health risk of the agent and animal housing density. b. Cage changing is performed twice a week for all static microisolator shoebox caging when housing rodents at standard densities. c. Cage changing for single housed mice in static microisolators may be performed at intervals longer than twice weekly, but should at least be performed once a week.

ABSL-2 Rodent Husbandry Effective 3/ Page 4 of 6

  1. The biological indicator is removed from the simulated “test” load and incubated in accordance with SOP #.
  2. The biohazardous load is securely stored in the dedicated locked freezer in IDRB room 428 until evidence that the biological indicator from the simulated load indicates that the biohazardous load has been successfully decontaminated.
  3. Results of monitoring are recorded on the Autoclave Sterilization Record.
  4. Autoclaved caging/equipment can only be removed from the locked freezer in IDRB room 428 and taken to IDRB cage wash room 406 for sanitation after efficacy of decontamination has been confirmed.
  5. Work surfaces are decontaminated after each use with bleach solution (1: dilution), followed by 70% alcohol rinse. Alternatively, an accelerated hydrogen peroxide (i.e., Oxivir Tb) can be used.

18. Protective clothing is removed and discarded in a red biohazardous container within the room/suite.

19. Hands are washed/sanitized prior to leaving the room/suite.

20. When biohazardous waste containers are to be removed/emptied containers/bags are sprayed with bleach solution (1:10 dilution), or Oxivir Tb and allow for appropriate contact time for disinfection (i.e., 5 minutes for Oxivir Tb and 2 minutes for bleach) prior to removing from the room.

  1. Room Duties a. Daily room duties: Upon completion of cage changing, feed and watering, and daily health surveillance, the following tasks are to be performed: (Room duties described below are minimal requirements, additional duties and frequencies are at the Facility Manager’s discretion). 1. Complete Room Status Sheet by recording: a. Minimum and maximum room or IVC rack temperature and humidity (measurements out of the acceptable range are described under the Health and Environmental Concern column of the Room Status sheet and reported to the Facility Manager via the Health and Environmental Concern Form ). b. Feed and water are available. c. Ventilation trolleys are functional (for IVC caging) d. Caging and equipment changes performed. e. Housekeeping duties performed. f. Significant health or environmental concerns. g. Time of observation and the initials of technician. 2. Clean/disinfect floor with Decon-Spore 200 Plus using a mop handle with a disposable cleaning pad/head. 3. Assure no investigator’s supplies or trash is left in the room. 4. Check and replenish supplies (e.g., soap, paper towels) as needed. 5. Wipe down counter/sink areas with Oxivir Tb 6. Assure any new arrivals are recorded on appropriate Per Diem Sheets.

ABSL-2 Rodent Husbandry Effective 3/ Page 5 of 6

  1. Record any additions or subtractions made by room technician or research staff as observed (e.g., weanlings, breeding pairs set up/removed, euthanasia, etc.) b. Weekly room duties include:
  2. Clean and disinfect changing station, if present, and clean/replace filters in accordance with SOP #1129 entitled NuAire Small Animal Cage Changing Station.
  3. Clean and disinfect biosafety cabinets, if present, in accordance with SOP #1127 entitled Biological Safety Cabinets.
  4. Wipe down door and door-frame with Oxivir Tb.
  5. Physically count animals within the room. c. Semi-monthly room duties include:
  6. Sanitize all room cleaning equipment within the room with, Oxivir Tb (e.g. wipe down mop handle, dust pan, etc.). d. Monthly room duties include:
  7. Clean HVAC vents.
  8. Confirm diurnal light timers are accurately controlling animal room lights SOP 1145 HOBO Data Logger , and record. e. Room surfaces and equipment are disinfected at the end of a study.

22. Animal Relocation Procedures a. Whenever rodents are to be relocated for use of common procedural areas outside of the ABSL-2 housing room (e.g., small animal imaging core, multiphoton microscopy core):

  1. Relocation is scheduled by Comparative Medicine staff and accomplished when no other animals or activities are present in the immediate area.
  2. Relocation is accomplished in clean primary enclosures, the exterior surface of which is decontaminated with Oxivir Tb prior to being removed from the housing room by Comparative Medicine staff.
  3. Surfaces and common equipment are decontaminated before and after use in accordance with SOP 1015 Decontamination of Common Procedural Areas.
  4. After return to ABSL-2 housing, primary enclosures used in transit are decontaminated by autoclaving.
  5. Complete cleaning and sanitization of the IVC racks at the IDRB facility requires that it be relocated to a facility with a rack-washer. Prior to removing the IVC rack from the BSL-2 suite the rack must be prepared for transport by following the decontamination and containment procedures below: a. While wearing appropriate PPE, remove all caging from rack being removed and place on the IVC replacement rack. b. Animals/cages will be moved to the new rack so that they retain the exact same cage position as they held on the original rack. c. Spray rack with Oxivir Tb thoroughly and allow for 5-minute contact time. d. Wrap rack with cellophane/plastic wrap thoroughly covering all surfaces so that rack is completely contained with only the wheels exposed. e. Spray wheels with Oxivir Tb as the rack is being removed from the room. f. Remove PPE prior to exiting the room. g. Roll rack to freight elevator and load onto vehicle for transport to rack-washer.