title: Evaluation of effects of high incubation temperatures on results of protozoal culture and real-time PCR testing for Tritrichomonas foetus inoculated in a commercially available self-contained culture media system.
abstract: OBJECTIVE
  To evaluate effects of high incubation temperatures on results of protozoal culture and real-time PCR testing for Tritrichomonas foetus inoculated in a commercially available self-contained culture media system.
  
  
  METHODS
  In vitro experimental study.
  
  
  METHODS
  2 strains of T foetus (1 field isolate from the University of California-Davis and 1 field isolate from the Texas Veterinary Medical Diagnostic Laboratory).
  
  
  METHODS
  2 sets of 36 dual-chamber media pouches were inoculated with T foetus (36 sample pouches/strain) and incubated at temperatures of 37.0°C (98.6°F), 46.1°C (115.0°F), or 54.4°C (130.0°F) for 1, 3, 6, or 24 hours. Six uninoculated media samples in pouches stored at 37.0°C for the entire treatment period were used as negative controls. Pouches were removed from incubators and stored at 22.2°C (72.0°F) until all treatments were complete. Samples were submitted to a diagnostic laboratory for protozoal culture and real-time PCR testing.
  
  
  RESULTS
  T foetus was detectable microscopically in inoculated pouches incubated at 37.0°C regardless of exposure time, whereas those incubated at 46.1°C yielded T foetus after 1 and 3 hours only, and those incubated at 54.4°C yielded T foetus after 1 hour only. Testing via real-time PCR assay yielded positive results for all inoculated media samples and negative results for all uninoculated control samples.
  
  
  CONCLUSIONS
  Samples collected into the self-contained culture media system for T foetus testing via culture alone should be protected from high temperatures. Realtime PCR amplification may be a more reliable method for identification of the organism if storage and transport temperatures cannot be controlled.
authors: Department of Large Animal Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX 77843, USA. john.davidson@novartis.com
authors: John M
authors: JM
authors: Davidson
authors: Jeff D
authors: JD
authors: Ondrak
authors: Arn A
authors: AA
authors: Anderson
authors: Amy K
authors: AK
authors: Swinford
authors: Erdal
authors: E
authors: Erol
mesh_terms: D000818:Animals; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis / Q000469:parasitology; D003470:Culture Media; D006358:Hot Temperature; D011529:Protozoan Infections, Animal / Q000175:diagnosis; D060888:Real-Time Polymerase Chain Reaction / Q000662:veterinary*; D016835:Tritrichomonas foetus / Q000502:physiology*
chemical_list: D003470:Culture Media