2010

SOP Number: CP042.00 Title: Procedure for Emptying Bladder in Catheterized Piglets Revision No: Replaces: 00 N/A Author: Kristin Smith I

Date in effect: Page: 7/15/2010 Page 1 of 5 MAL Director: Dr. Peggy Borum

FSHN Chair: Dr. Neil Shay

PURPOSE The purpose of emptying the bladder of catherized piglets is to collect urine for analysis during the experiment.

II

SCOPE This procedure provides instructions for emptying the bladder in catheterized piglets.

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 N/A

V

REAGENTS AND MATERIALS V.A. V.B. V.C. V.D. V.E. V.F.

VI

70% Ethanol. Kimwipes. 22-gauge needle. 3cc syringe. Labeled white-capped plastic scintillation vial. Ice (and bucket).

EQUIPMENT N/A

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.

VIII

DEFINITIONS

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SOP Number: CP042.00 Title: Procedure for Emptying Bladder in Catheterized Piglets Revision No: 00

Replaces: N/A

Date in effect:

7/15/2010

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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 Identification (ID)—The Piglet ID is a unique, alphabetical and numerical label that identifies the piglet and the samples taken from that piglet. IX

PROCEDURE IX.A. Pre-experiment. IX.A.1. Label sample vials to be used in the experiment in advance. IX.A.2. The following information should be marked on the urine vials: IX.A.2.a. Piglet ID. IX.A.2.b. Experiment day. IX.A.2.c. Time period to begin using that vial. IX.A.2.d. Vial number in case that more than one vial is used during the hour long period. IX.B. Inside the PNICU. IX.B.1. Open a 3cc syringe and attach a 22-gauge needle. Between uses, loosely recap syringe and place in a designated area (either on top of incubator or on a moving cart as shown in Figure 1). Always use a 3cc syringe, even if the volumes of urine become large.

Figure 1. IX.B.2. The urine syringe should be changed every 6 hours. IX.C. Inside the entry room. IX.C.1. Fill the ice buckets with ice. IX.C.2. Place appropriately labeled scintillation vials next to ice bucket. IX.C.3. Place the vials on ice that will be used within the hour. IX.D. Emptying the bladder catheter.  Uncontrolled Copy

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SOP Number: CP042.00 Title: Procedure for Emptying Bladder in Catheterized Piglets Revision No: 00

Replaces: N/A

Date in effect:

7/15/2010

IX.D.1.

IX.D.2. IX.D.3. IX.D.4.

IX.D.5. IX.D.6. IX.D.7. IX.D.8. IX.D.9. IX.D.10.

IX.D.11. IX.D.12. IX.D.13. IX.D.14.

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The catheterized bladders of piglets that have not had their urethra ligated should be emptied at 30 minute intervals. Two urine collections will be placed in the same vial to represent the urine collected during that hour. Swab the septum of the bladder catheter with a Kimwipe and 70% ethanol. Insert the needle of the syringe into the septum. Draw back on the syringe gently to aspirate any urine present in the bladder as shown in Figure 2.

Figure 2. IX.D.4.a. The same needle and syringe can be used for one piglet for 6 hours before being replaced. IX.D.4.b. When not in use, the syringe, loosely capped, should be kept on top of the piglet's incubator. Slowly draw urine until you feel a change in pressure. Do not pull too hard on the syringe as this can cause damage to the bladder. Stop if a large amount of tissue appears in the syringe or if is difficult to pull the plunger. Note the amount and quality of urine. Note the total amount, color and clarity of the urine on the piglet's medical record. If no urine is collected, record this as 0mL on the chart. If the collected urine is markedly different from the previous collection, contains blood, clots, clumps, or is of an odd color, put the urine into a separate vial. Notify Dr. Borum of the abnormal urine. Label the vial as indicated in SOP# CP035.00 When the urine collected returns to normal, resume using the regular prelabeled vials. Deposit the urine in the appropriately labeled vial as shown in Figure 3.

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SOP Number: CP042.00 Title: Procedure for Emptying Bladder in Catheterized Piglets Revision No: 00

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Date in effect:

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Figure 3. IX.D.15. Do not fill the urine vials more than 75% full if they are to be frozen to prevent bursting of the vials. IX.D.16. The same vial can be used for every two urine draws (two draws per hour). IX.D.17. These urine vials should be kept in the ice bucket inside the PNICU’s entry room workspace and/or cabinet until they are transferred to the lab for analysis. Analysis will be performed only after urine for that hour has been collected (See SOP# CP020.00). IX.D.18. Confirm that the screw caps are tightly fastened before placing the vials on ice. IX.D.19. Periodically, drain the ice bucket of the water that forms and refill it with fresh ice. IX.D.20. Be sure to keep track of samples and piglets so that each sample goes into the correct vial.

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SOP Number: CP042.00 Title: Procedure for Emptying Bladder in Catheterized Piglets Revision No: 00

X

Replaces: N/A

Date in effect:

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ATTACHMENTS X.A. For a list of materials and their locations, refer to SOP# CG098.00.

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