Page: Page 1 of 9 Author: Kristin Smith MAL Director: Dr. Peggy Borum FSHN Chair: Dr. Neil Shay

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1 Page 1 of 9 Author: Kristin Smith MAL Director: Dr. Peggy Borum FSHN Chair: Dr. Neil Shay I PURPOSE Incubators serve to maintain stable temperature and humidity levels suitable to the survival and health of the piglet. II SCOPE This procedure provides instructions for the preparation of piglet incubators before an experiment. The incubators need to be clean and maintained in an aseptic environment for the experiment. III RESPONSIBILITIES It is the responsibility of Metabolic Assessment Laboratory personnel to follow this procedure. It is the responsibility of supervisory personnel to ensure compliance with this procedure and to train employees and students responsible for performing this procedure. Students will report accidents to the principal investigators immediately. IV REFERENCES V REAGENTS AND MATERIALS V.A. Disinfectant cleaning solution of a mixture of 4 oz of disinfectant to 2 gallons of water is used to clean incubator. V.B. Autoclaved piglet floors. VI EQUIPMENT VI.A. The piglet incubator is made of plexiglass with an inside base of inches in length, 14.5 inches in width, and walls that are 13.5 inches in height. A 10 x 26 inch hinged front door provides easy access to incubator. VI.B. The piglet incubator solid door(s) can be replaced with a 10x26 inch bar grid door, which is hinged to provide easy access to incubator. VII SAFETY PRECAUTIONS VII.A. Members of the MAL have been trained extensively in the procedures described in this SOP. VII.B. Members of the MAL have completed animal contact training in order to work in the PNICU.

2 2 of 9 VIII DEFINITIONS VIII.A. Standard Operating Procedure (SOP) Standard Operating Procedure is a document that provides instructions for completing a specific task in the lab. VIII.B. Metabolic Assessment Laboratory (MAL) The Metabolic Assessment Laboratory is the laboratory that will use this SOP. VIII.C. Piglet Neonatal Intensive Care Unit (PNICU) The PNICU is a unit where the piglet is monitored by 24 hour care and routine check-up parameters using PNICU SOPs conducted by the MAL. VIII.D. Piglet Incubator The Piglet Incubator is an infant incubator that is enclosed, lined with piglet floors, and temperature and humidity regulated. Piglet Incubator

3 3 of 9 VIII.E. Incubator Bedding Tray The bedding tray is the bottom of the incubator where the piglet and other items may be placed. See picture below for description. Incubator Bedding Tray VIII.F. Incubator Deck Rest The Hood Assembly rests on a deck gasket that fits on the outer limits of the deck rest. The Hood Assembly must at all times rest on the deck gasket. See picture below for description. Deck Rest

4 4 of 9 VIII.G. Incubator Deck Gasket The deck gasket fits on the outer limits of the deck rest. See picture below for description. Deck Gasket VIII.H. Incubator Micro Oxygen Filter The Micro Oxygen Filter filters all air drawn into the piglet incubator. The Micro Oxygen Filter must be checked before each experiment. Clean or change as needed. See picture below for description. Micro Oxygen Filter

5 5 of 9 VIII.I. Incubator Power Pack The control power unit uses a heater to warm the piglet. Temperature may be controlled manually using the air temperature control knob. See picture below for description. Power Pack VIII.J. Incubator Humidity Fill Tube The Humidity Fill Tube is used to pour water into the humidity reservoir. It also serves as an indicator as to the level of water in reservoir. To drain water from reservoir, gently turn Humidity Fill Tube upside down. See picture below for description. Humidity Fill Tube

6 6 of 9 VIII.K. Incubator Humidity Reservoir Tank and Baffle The Humidity Reservoir Tank holds water to provide proper humidity in the incubator. A Baffle is used to stabilize water in tank reservoir. See picture below for description. Humidity Reservoir Tank and Baffle VIII.L. Incubator Humidity Control Knob The Humidity Control Knob on the left side of the Hood Assembly controls the level of humidity inside the incubator. The knob must be pushed in to come into contact with control place located on deck rest. See picture below for description. Humidity Valve (Control Plate)

7 7 of 9 VIII.M. Incubator Entry Port Door(s) The Entry Port Door(s) provide arm access to the incubator from the front, side, and rear. Doors are latched to provide easy entry/exit. One larger hinged entry door on front provides greater access. See picture below for description. Entry Port Doors VIII.N. Piglet Floor The Piglet Floor is 7-8 sheets of pre-cut newspaper wrapped in butcher paper and then sterilized to line the floor of the incubator. VIII.O. Piglet Toys Piglet Toys include balls that have been disinfected for piglet play. VIII.P. Teddies Teddies are sterilized towels that serve as blankets for the piglets. IX PROCEDURE IX.A. Preparation of Incubator. IX.A.1. Before cleaning PNICU piglet incubator, unplug the power cord from wall receptacle and remove micro air filter. Note: Filter must not get wet. IX.A.2. Use handle on back of plexiglass hood to lift hood forward. IX.A.3. Remove incubator bedding tray. IX.A.4. Lift the deck rest and remove (deck must never rest on deck gasket). IX.A.5. Wash the plexiglass incubators. IX.A.5.a. Wash with disinfectant cleaning solution. Use a soft cloth to clean because the plexiglass will scratch very easily. IX.A.5.b. Rinse the plexiglass incubators with tap water. IX.A.5.c. Never use alcohol on the incubators because it could result in cracks or fissures to the plexiglass. IX.A.5.d. Clean piglet incubator humidifier air reservoir with disinfectant cleaning solution.

8 8 of 9 IX.A.5.e. Check the micro oxygen filter for cleanliness and change if necessary. IX.B. Clean and check power pack. IX.B.1. Unplug the power cord from the power pack on the right side of incubator, unscrew knobs from front panel, and pull entire unit out using great care. IX.B.2. The heater must not be touched until it has cooled for at least 20 minutes. IX.B.3. Remove lid from heater coil and clean blades with air. IX.B.4. Remove oxygen intake tube (turn and pull forward) and clean with disinfectant and a clean cloth. IX.C. Clean the humidity fill tube located on left front below hood. IX.C.1. Unscrew the humidity fill tube and clean tube. Note: Use caution because they are easily broken. IX.D. Clean the humidity reservoir tank and baffle with disinfectant cleaner and soft cloth. IX.D.1. Remove the humidity valve (control plate) from left inside hood assembly by removing two retaining thumbscrews. Clean with disinfectant cleaning solution and soft cloth. IX.D.2. Replace port sleeves on left side of incubator. IX.D.3. Carefully remove entry port assembly retaining rings on front of and back of incubator and clean with a disinfectant cleaning solution. IX.D.4. Place a squirt bottle of saline and 70% alcohol, and box of KimWipes on top shelf of incubator, for use during 24-hour care. IX.D.5. Place two autoclaved piglet floors on the bottom of each incubator so that they overlap,f or use during 24-hour care. IX.D.6. Put an autoclaved teddie (towel or linen) in each incubator, for use during 24-hour care. IX.D.7. If needed, set a baby bottle nipple (that has been autoclaved in water) in the bottle holder on the outside of the incubator if available or tape the nipple to the rear inside wall of the incubator to be used as a pacifier, for use during 24-hour care. IX.D.8. Place a wiffle ball that has been soaking in a disinfectant solution on the piglet floor, for use during 24-hour care. IX.E. Preparation of Cabinets on which the Incubator rests. IX.E.1. Wash the painted base cabinets with disinfectant solution. IX.E.2. Clean the plexiglass medical record holder with disinfectant solution. Use a soft cloth to clean because plexiglass will scratch very easily. Place the clean plexiglass medical record holder on the side of the cleaned incubator. IX.E.3. Clean the infusion pump if needed and place to the side of the incubator. IX.E.4. Place the medical record in the plexiglass medical record holder. IX.E.5. Tape an identification tag to the plexiglass medical record holder of each piglet. IX.E.6. Place one package of autoclaved piglet floors, one package of autoclaved teddies, one box of KimWipes, and one roll of paper towels into the cabinet under the incubator.

9 9 of 9 X ATTACHMENTS X.A. For a list of materials and their locations, refer to SOP# CG098..