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Kerry Rood: Trichomoniasis caused by the protozoan tritrichomonas foetus is a
serious reproductive or venereal disease found in some Utah cattle herds.
Diagnosis of trichomoniasis is made when the trichomonad organism are detected
in the smegma or preputial flush samples of bulls or the uterine vaginal fluids
of cattle.
Because of the potential contamination of the sample with fecal trichomonads,
the use of an approved polymerase chain reaction, or PCR test differentiates
between the two.
Frozen samples are to be shipped on ice to the Utah Veterinary Diagnostic
Laboratory. [Folksy string music]
Rood: Such that arrival can be guaranteed to happen when the laboratory's open
and the sample remains frozen, we suggest overnight delivery.
Once frozen, the sample can remain in the freezer until delivery is arranged.
Call the Utah Veterinary Diagnostic Lab at 435-797-1895 to ensure delivery and
to alert them of incoming samples.
A sample that is inconclusive will be considered positive.
A sample determined to be negative to tritrichomonas foetus by PCR will be
considered negative.
Sample collection. With bulls, the preferred sample location at the
glans penis. The glans penis is the area between the sheath and penis.
Any hanging fecal balls or long hair at the preputial opening should be trimmed
to minimize sample contamination.
Samples are obtained using a sterile insemination pipette and performing a
vigorous back and forth scraping motion along the glans while applying negative
pressure with an attached 10 or 20 mil syringe.
The manufacturer's media package insert recommends a 20 mil syringe be used.
Keep in mind a separate sterile pipette and sterile syringe should be used for
each animal.
After sampling, a current official trich tag should be placed in the right ear of
each bull at the time of sample collection under the direction of a
certified veterinarian. Official trich tags can be obtained by contacting the
Utah Department of Agriculture and Food. [Folksy string music]
Rood: The color of tag changes each year and the markings correspond to the year
and to the state.
Media. Currently, the only official recognized media for the culturing of
bovine trichomoniasis organisms in the state of Utah is the InPouch,
manufactured by Biomed Diagnostics.
For media inoculation, hold the pouch upright, tear off the upper edge where
marked to allow access to the upper chamber. If some fluid from the lower
chamber is not in the upper chamber, gently apply pressure to the lower chamber
to transfer media from bottom to top.
Inoculate the sample into the small upper chamber of the pouch.
Flush out the pipette by drawing up a few milliliters of pouch solution into the
pipette, repeating as necessary to sufficiently inoculate the upper chamber.
The inoculated fluid in the upper chamber should be squeezed or squeegeed into the
lower chamber, being careful to apply just enough pressure to evacuate the upper
chamber without rupturing the pouch itself.
Carefully express air bubbles out of the lower chamber to maintain the anaerobic
environment. Roll the top of the plastic pouch down to the top of the lower chamber
and fold the wire strips across to hold and seal.
Caution: do not stir or mix the contents and keep the packet upright at all times
once the lower chamber is inoculated.
Handling and care of the pouch is as critical. The handling and/or shipping of
the inoculated media samples is one of the most critical steps in trichomoniasis
diagnosis. The inoculated media should be kept at 65 to 90 degrees Fahrenheit
until it is incubated. It is especially important to avoid overheating or
freezing. Handle the inoculated pouches in insulated containers that will protect
the samples from extreme temperatures, either hot or cold.
Trichomonad organisms are very susceptible to either freezing or overheating.
Samples should be incubated as soon as possible at 98 degrees Fahrenheit or
37 degrees Celsius for 24 hours.
After incubation, pouches should be frozen directly. Frozen samples should be shipped
in an insulated container to ensure samples remain frozen.
It is important to arrange shipping so that samples arrive at the laboratory
in a frozen state.
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