Blood sampling
From rodent captured, 1 to 2 ml of blood was aseptically collected from the heart puncture or plexus by using sterile syringes and needles. In cattle, which were randomly sampled from different herds, 4 to 10 ml of blood was aseptically collected from the jugular using sterile syringes and needles after proper manual animal restraint. In humans, the study involved individuals with febrile illness who were visiting Teule Hospital and Ubwari Health Center. A 2 to 4-ml blood sample was aseptically collected from each study subject by medical personnel using sterile syringes (Mgode et al., 2019). Socio-demographic information of study participants, including sex, occupation, location, and age was recorded.
The samples were immediately transferred into plain vacutainer tubes and allowed to clot for serum separation at room temperature for at least 30 minutes. The samples were centrifuged for 10 minutes at 3000 rpm to increase the serum volume. The harvested sera were subsequently transferred into well-labeled Eppendorf tubes and then stored at -20ᵒC until subjected to MAT. (Mgode et al ., 2019).