Tritrichomonas foetus PCR Testing

Storage and shelf life of In-Pouches and Transit Tubes

Do not refrigerate or freeze. Store at room temperature, vertically, way from direct light. Do not use expired media. Do not use if the liquid appears to be cloudy, leaking, dark brown or dried.

If you are in need of supplies, please contact the NVDC at 402-472-8470.

Sample preparation

Clip the hair around the preputial orifice in bulls. Flush the preputial cavity with sterile saline solution (do not use water) to clean out the mud and manure if necessary. This process will help to decrease the risk of overgrowth of non-T. foetus bacterium).

Sample collection

Bulls: Direct the pipette to the distal penis in the sheath Scrape the mucosa of the distal penis and the fornix area while applying suction with syringe or bulb to obtain the specimen.

Cows: Advance the pipette gently to the floor of the vaginal fornix, and aspirate mucus.

Preparation of In-pouch TF or TF Transit Tube

Label the In-pouch/Transit Tube with the sample related information.

Samples for PCR should be submitted in a TF Transit Tube. If samples are submitted in InPouch TF media, a processing fee will be applied.

TF Transit Tube: Open the transit tube by twisting off the cap. Be sure when removing the cap that you are ready to use the tube as doing so will break the seal.

InPouch TF: Manually express the liquid so all of the liquid is in the lower chamber. Open the pouch by tearing off the top. There is a preformed score to facilitate tearing. Use the integral white tabs to open and secure the mouth of the InPouch TF open.

Inoculation of sample

Insert 0.5-1.0 ml of specimen into the TF Transit Tube/InPouch TF. Secure the cap on the tube or express the entire contents of the InPouch TF into the lower chamber. If using an InPouch TF, avoid trapping air. Roll the InPouch TF top tightly until the wire-tape is at the top of the label and fold the wire tabs to seal the pouch.

Incubation and shipping of samples

T. foetus samples submitted in TF Transit Tubes for PCR do not require incubation. After collection, store TF Transit Tubes in either a refrigerator or freezer, in the dark. Ship samples on ice.

T. foetus samples submitted in InPouch TF media for PCR must be incubated before testing. If you wish to incubate prior to shipping, please incubate the samples in the dark, vertically, at 35-37°C for 24 hours. If you do not have an incubator, a Styrofoam container with an inserted microwaved ice pack (warm to the touch) or hand warmer can be used to create a makeshift incubator. Do not place the InPouch TF or TF Transit Tubes directly on the heat source as they may overheat. If incubation is performed by the submitting veterinarian, please note this on the submission form.

• Ship InPouch TF samples not yet incubated overnight at room temperature without ice. If outside temperatures are exceedingly warm, ice packs may be placed inside the shipping container. If ice packs are included, do not allow the ice to come into direct contact with the samples or allow the InPouch TF media to freeze.
• Samples in InPouch TF media that have been incubated prior to shipping can either be sent with ice or at room temperature.

Additional instructions and guidelines

All samples must be inoculated into unexpired media and receoved within 5 days of collection unless frozen.

If the samples require pooling, the submitting veterinarian MUST note this on the submission form. The UNL NVDC cannot pool T. foetus samples at their own discretion. States vary in their acceptance of pooling for import requirements. Please check with the state animal health official for their import requirements.

Samples cannot be pre-pooled by the submitting veterinarian. This must be performed only by the diagnostic laboratory.