vivax genome compared to T. congolense. Currently, the 18S based PCR-RFLP is the FAO recommended test for detection and differentiation of multiple trypanosome taxa ( Geysen et al., 2003). The 18S PCR-RFLP requires one or two amplification

steps and a restriction digestion step. On the other hand, the ITS1 TD PCR needs a single amplification step and achieves a comparable detection limit, is thus less-time consuming and more cost-effective for detection of T. congolense, T. vivax and the three species within the Trypanozoon subgenus E7080 T. brucei, T. evansi and T. equiperdum. The data obtained with the blood specimens of experimentally infected cattle demonstrated that ITS1 TD PCR had comparable performance to microscopy in revealing the absence or presence of

a T. congolense infection prior to treatment. After curative treatment, the ITS1 TD PCR became negative in all animals within no more than two days, thus proving the rapid clearance of DNA from dead parasites in the blood. The discordant result at day 1 post-treatment in animal CVB 190, with positive HCT and negative ITS1 TD PCR was confirmed with a negative result in the 18S-based real-time PCR developed in our laboratory (unpublished), but cannot be explained. In the animals that relapsed after treatment, ITS1 TD PCR achieved a higher positivity rate and was able to detect recrudescence of parasitaemia several days earlier than HCT. Therefore, ITS1 TD PCR can be considered Ferroptosis inhibitor review more reliable than microscopy in assessment of trypanocide efficacy and bears the potential to allow shortening of the post-treatment follow-up that is currently recommended to be 100 days. In summary, the ITS1 TD PCR offers an alternative tool for trypanosome

diagnosis and assessment of compound efficacy against T. congolense in cattle. The ability to detect and differentiate multiple Trypanosoma taxa in a single run, with the potential to identify mixed infections, indicates the application of the ITS1 TD PCR for research on other pathogenic trypanosomes, such as T. vivax, T. evansi, T. equiperdum and T. brucei. None of the authors has of any financial or personal relationship that could inappropriately influence or bias the content of the paper. This research was supported by the Global Alliance for Livestock Veterinary Medicines (GALVmed) with funding from the UK Government’s Department for International Development (DFID) as part of GALVmed’s Animal African Trypanosomosis Program (DFID Programme: Controlling African Animal Trypanosomosis (AAT) (Aries code 202040-101). The authors are grateful to Dr. Filip Claes for the in silico work. We thank Fatima Balharbi, Nicolas Bebronne and Stijn Rogé for technical assistance. For the veterinary clinical trials (animal husbandry, veterinary supervision, project management and data management), we acknowledge the ClinVet International (Pty) Ltd staff. T.