Addressing the constraints of Tritrichomonas foetus sample collection in remote areas: lyophilized modified Diamond's media as a substitute for liquid medium.

Bovine trichomoniasis is a notifiable, reproductive disease of cattle caused by the parasite Tritrichomonas foetus. Culturing with modified Diamond's medium (MDM) is required to increase the low number of organisms received from a preputial sample, but is limited in application to remote areas as it requires continuous cold chain storage. This study utilized lyophilization to sustain the viability of MDM during transport in lieu of a continuous cold chain. All lyophilized MDM was able to sustain T. foetus after storage for 42 days at 24 °C, and the results demonstrated that lyophilized MDM was equally as viable as refrigerated liquid MDM. Storage of lyophilized MDM at room temperature for 1 and 7 days did not impact T. foetus yield, both with and without exposure to light. A limitation of the lyophilized MDM was demonstrated with a significant decrease in T. foetus yield when the media was stored at 37 and 58 °C. The lyophilization of MDM provides a robust method of transporting and storing medium prior to reconstitution and inoculation, for use in T. foetus diagnosis and surveillance in remote areas.

Evidence of self-resolution of feline trichomonosis in a pair of single household cats due to ronidazole-resistant Tritrichomonas foetus.

Tritrichomonas foetus causes chronic large-bowel diarrhoea in cats, complicated not only by its chronicity but for its difficulty to treat, with few registered or effective products available, subsequently resulting in ronidazole frequently used off-label. The extent of T. foetus resistance to ronidazole is unknown. To demonstrate in vitro resistance of feline T. foetus to ronidazole and the self-resolution of the infection, we isolated novel T. foetus "feline" genotype strains from two cats in 2017 from the same household and monitored these cats over a three-year period. Axenic cultures of T. foetus from the cats were tested for in vitro resistance to metronidazole and ronidazole using anaerobic and aerobic culture assays. The minimum lethal concentration for the novel strains after incubation in aerobic conditions for 48 h were 50-100 µg/ml and 6.25-12.50 µg/ml for metronidazole and ronidazole, respectively. For susceptible T. foetus strains, the minimum lethal concentrations ranged from 1.56-12.50 µg/ml for metronidazole, and 0.39-3.13 µg/ml for ronidazole. The self-resolution of the infection was assessed using repeat diagnostic qPCR and culture. Whilst positive in 2018, the cat samples are no longer returning positive qPCR results in 2020 and 2021, indicating self-resolution over this period. This study demonstrated resistance to ronidazole for the first time in Australian cats and infection self-resolution without antimicrobial intervention is demonstrated. This study provides clinicians with the evidence to reduce the use of off-label ronidazole, and advice on the resolution timeframe for cats in single households.