California licensed, USDA accredited veterinarians can request a training video to become approved for trichomonosis sample collection in California through their Animal Health Branch District Office. This DVD will be mailed to the requesting veterinarian to watch independently; it will arrive with a form that must be returned to the district office once the veterinarian has watched the training video. A certificate of training for sampling and handling will be mailed from AHB headquarters following receipt of this form at the district.

Once the form has been received by CDFA, you are approved for sampling and handling of trichomonosis samples. Approval is renewed every two years with CDFA at the same time Brucellosis contracts are renewed.

Clinics can become approved to read trichomonosis samples by sending someone from the clinic to a training session at a California Animal Health & Food Safety (CAHFS) Laboratory. This person may be a veterinarian or another clinic employee designated by the clinic / laboratory.

Multiple individuals from a clinic are welcome to attend training sessions; however, please inform CDFA in advance of the names and job functions (veterinarian, technician, RVT) of all people planning to attend so the lab can plan accordingly. Each CAHFS lab training session lasts about one hour, and is limited to no more than six (6) people.

The expectation with lab training is that the person trained at CAHFS will go back to the clinic and share knowledge gained with all individuals reading trichomonosis samples.

If the person attending this lab training subsequently leaves the clinic, then that clinic/lab is no longer approved to read trichomonosis samples. If this happens, please contact CDFA to schedule another person for lab training.

Scheduling can be done online at: http://et.cdfa.ca.gov/trich/selfreg/. If veterinarians / clinics do not have web access, they can contact their Animal Health Branch District Office to complete the online scheduling form. CDFA will then contact the individual to confirm training date and location.

All positive tests must be reported to your local Animal Health Branch District Office within two (2) days of the final read date.

All negative tests must be reported to your local Animal Health Branch District Office within 30 days of the final read date.

All test results should be reported on CDFA’s Trichomonosis Test Report Form. If a CAHFS laboratory reads the samples, they will report results to the Animal Health Branch.

Bull owners or their testing veterinarian may request PCR confirmation of a positive culture result. If confirmation by PCR has been requested, your Animal Health Branch District Office should be informed of the positive culture result, but the positive test result will not be investigated until the PCR result is reported. For the positive culture result to be reclassified as a negative official test, the PCR must be negative for T. foetus and positive for other trichomonads. CAHFS laboratories will report final test results directly to CDFA, so the testing veterinarian will not need to do so.

The laboratory will report PCR results as follows:

• If the bull is PCR positive, using primers specific for T. foetus , he is declared infected.
• If the bull is positive for trichomonads in general, but negative for Tritrichomonas foetus, the bull is reported as negative.
• If the PCR is negative for all trichomonads, it suggests that there was an insufficient amount of DNA present to be detected by the PCR assay or that the DNA was somehow degraded, and a re–test is recommended.
• In very rare cases, in spite of a live culture being submitted for testing, PCR tests may be equivocal, in which case a re–test should be done.

The most commonly used culture medium, the InPouch TF, is available from BioMed Diagnostics. BioMed also sells a T. foetus live culture (positive control); this is an excellent reference sample for clinics to compare to samples collected from client animals.

Prevent contamination during sample collection. Bacteria grow easily in the culture medium.

Ways to minimize bacterial overgrowth of samples:

• Use a new pipette for each collection
• Use pipettes covered in a soft plastic chemise; pop end of pipette through chemise only once it is in the proper location for sampling
• Trim preputial hair if long or dirty
• Flush prepuce with 60 – 120cc sterile, non–bacteriostatic saline if there is gross contamination such as mud or manure
• Re–sample if chunks of manure or debris are seen in culture pouch after inoculating