STANDARD OPERATING PROCEDURES

DIVISON OF COMPARATIVE MEDICINE

UNIVERSITY OF SOUTH FLORIDA

SOP#: 1011.1

Date Issued: 12/03

TITLE:
SCOPE:
RESPONSIBILITY:
PURPOSE:

I.

Date Revised: 1/12

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RODAC Plate Procedures

Animal Care Personnel
Facility Manager and Technical Staff
To Outline the Proper Procedures for Obtaining, Processing and
Recording Results of RODAC Samples for Sanitation Monitoring

PURPOSE
1. RODAC (Replicate Organism Detection And Counting) plates are used in
conjunction with the novaLUM portable analyzer and PocketSwab Plus (See SOP
#1010 Microbiological Monitoring of Sanitation Procedures and SOP #1014
Monitoring Animal Drinking Water)) to assess the efficacy of cleaning and
sanitation procedures, and assuring the quality of potable water for research animals
as part of the overall microbiological monitoring program.

II.

RESPONSIBILITY
1. Facility Managers are responsible for ensuring that tests are performed properly.
Managers will ensure that plates and swabs are marked with proper codes that
correspond to the Microbiological Monitoring Sample Log and are transported to
the College of Medicine (COM) facility on the same day samples are taken.
Managers may designate a staff member to perform testing and transportation.
Managers will obtain plates and sterile swabs on an as needed basis.
2. The COM Facility Manager or designated staff member is responsible for receiving
samples, incubating RODAC plates, plate counts, performing gram stains, and
recording all results. The COM Facility Manager will notify the Facility Manager
requesting testing of any marginal or unacceptable results by E-mail upon obtaining
results.

III.

PROCEDURE
1. RODAC plates are used to assess the efficacy of sanitation procedures after two
consecutive novaLUM test failures. Unlike the novaLUM, RODAC plates can
detect the presence or absence of live microorganisms.
2. RODAC plates must be stored refrigerated at 40 F with the topside up (agar bed at
bottom). Do not freeze. Do not open until ready to use. Minimize exposure to light.
Sterile culturette swabs may be stored at room temperature. Always use gloves
when handling plates and culturette swabs. Plates and swabs may be used up to
their expiration date. Discard expired plates and swabs.

SOP #1011.1
RODAC Plate Procedures
Effective 1/12
Page 2 of 4

3. Caging, walls, and procedural areas will be tested using RODAC plates directly.
Surfaces not conducive to direct plating (e.g., water bottles sipper tubes, wire
feeders, and water samples will be tested using sterile culturette swabs and
subsequent plate inoculation. Plastic, glass, and metal surfaces must be cool and
dry before sampling. When testing room walls, using RODAC plates allow the Clidox
to dry and dissipate before sampling.
4. Testing will only be performed on Mondays, Tuesdays, and Wednesdays. Samples
taken on Wednesday must arrive at the College of Medicine facility no later than
12:00 noon to complete incubation by Friday.
5. The Microbiological Monitoring Sample Log is initiated at each facility at the time
of sampling. The Facility Manager ensures that the Site, Date and Time Sampled,
Sampled By, Reason for Sampling (RT: Retest), Sample Code and Room Number
or Source are completed. Sample Code and Number must be assigned to a specific
item or room on the Microbiological Monitoring Sample Log prior to sampling the
item or room. The Remarks section of the log should indicate the date and time of
the failed novaLUM test.
6. Sample Codes are as follows:
RM Room Wall
MC Mouse Cage
ST Sipper Tube
WB Water Bottle

RK Rack`
RC Rat Cage
WF Wire Feeder
NHP Primate Cage

CG Cat Cage
GP Guinea Pig Cage
RB Rabbit Cage
WS Water Source

7. Each sample accompanying the Log must have full date and time of collection, site,
source, Sample Code legibly recorded on side of RODAC plate or culturette swab
label (i.e., 1/15/12 3:52 PM COM 1341 MC). Mark the RODAC plates on the outside
edge of the lower plate containing the agar bed with a fine point permanent marker.
Label culturette swabs using an appropriate label or tape.
8. The Manager or designee will complete the remainder of the Microbiological
Monitoring Sample Log. When the Microbiological Monitoring Sample Log is
complete the sample log will be sent to the Facility Manager requesting testing and
the Microbiological Monitoring Program Manager.
9. RODAC plates are specially prepared so that the agar forms a convex or domed
surface that can be pressed onto a flat surface for sampling. The plates are shipped
in a double bag. The inner bag and plates are sterile. Prior to use the plates must
be warmed to room temperature (15-20 minutes) in the original packaging. After
warming, carefully open the outer bag using gloves in a clean, closed room.
10. Carefully remove and open the inner bag. Remove the number of plates required for
sampling and reseal the inner and outer bags with tape. Write the sample numbers
on the outside edge of the lower plate containing the agar bed with a fine point
permanent marker.
11. Carry the plates in a small Styrofoam or other appropriate container to the cage or
cage rack drying area or to a disinfected room. Cages and cage racks must be cool
and dry before sampling. Room walls must be dry. Wearing gloves remove the

SOP #1011.1
RODAC Plate Procedures
Effective 1/12
Page 3 of 4

plate (top and bottom) from the container. Using one hand remove the top of the
plate. Hold the bottom of the plate with thumb and second finger and use index finger
to press the agar surface firmly against the test surface for five seconds. Pressure
should be the same for every sample. Make sure that the entire agar surface
contacts the test surface. Do not move the plate laterally as this spreads the
contaminants over the agar surface making resolution of colonies difficult.
Immediately after exposure, replace the top, tape the plate closed, and put into the
container. Record the date/time samples were taken on the Microbiological
Monitoring Sample Log.

12. Water bottles, sipper tubes, and wire feeders are sampled using sterile culturette
swabs. Bottles, sipper tubes, and wire feeders must be cool and dry before
sampling. Wearing gloves carefully open and remove the outer packaging of the
swab. Remove the top of the culturette with the cotton swab, moisten the swab with
sterile saline, and wipe the inside of the bottles and sippers and the metal surfaces of
the feeders. Immediately after exposure, replace the top and put into the container
with the RODAC plates. Record the date/time samples were taken on the
Microbiological Monitoring Sample Log.
13. On receipt at the COM facility, the Manager or designee will record the date and time
received on the Microbiological Monitoring Sample Log. The culturette swabs are
streaked onto RODAC agar plates. All plates are incubated at 35 -37 C for 48
hours. When placing the plates on the incubator shelves record the shelf number on
the sampling log to facilitate locating the plates after incubation.
14. The plates are checked for growth after 48 hours and the number of colony forming
units (CFU) are counted and recorded on the Microbiological Monitoring Sample
Log. High colony counts (>20) should be described in the comments column (e.g.,
all one type, small, diffuse, etc.). Gram stains are made of representatives of all
colonies on the plates and the results are recorded on the Microbiological
Monitoring Sample Log. The presence of Gm+ rods or Gm- cocci will be annotated
in the Remarks column.
15. The following scales are used to evaluate the sanitation process:
a. Colony Counts:
Excellent Sanitation = 0 CFU
Adequate Sanitation = 1-10 CFU
Marginal Sanitation = 11-20 CFU
Unsatisfactory Sanitation = >20 CFU
b. Gram Stain Results:
Excellent Sanitation = no growth
Adequate Sanitation = Free of gram negative rods and gram positive cocci
Marginal Sanitation = Free of gram negative rods, but some
gram positive cocci
Unsatisfactory Sanitation = Sample contains gram negative rods
16. Facility Managers will be notified of marginal and unsatisfactory microbiological
monitoring results within 48 hours. Facility Managers are responsible for
investigating and correcting the cause of marginal and unsatisfactory results per
SOP #1010 Microbiological Monitoring of Sanitation Procedures.

SOP #1011.1
RODAC Plate Procedures
Effective 1/12
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17. Used plates and swabs are disposed of as biohazardous waste.

IV.

EQUIPMENT AND SUPPLIES

1. RODAC plates using trypticase soy agar with lecithin and polysorbate 80. The
lecithin and polysorbate 80 are neutralizers to inactivate residual disinfectants. The
agar surface is dome shaped to allow the entire surface of the media to be pressed
against flat surfaces.
2. Culturette swabs.
3. Gram Stain Kit.

Approved:

Date: