A total of 45 Trypanosoma congolense

strains were isolated from communal cattle (Ngoni breed) reared in a trypanosomiasis endemic area located in the Katete and Mambwe Districts of the plateau areas of eastern Zambia (9). The area is highly cultivated with a cattle population of approximately 8–10 animals/km2. Cattle constitute the main host of the tsetse flies and are the main reservoir of trypanosomes (11). Large game animals are absent. Another five T. congolense strains were also isolated from communal cattle (Ngoni breed) kept in the Siyavonga District in the Southern Province of Zambia. The area is separated from the tsetse-infested wildlife area between Chirundu and Kariba in Zimbabwe by the Zambezi River. In both areas, cattle click here infected with T. congolense were identified BVD-523 using parasitological diagnostic tests (12). For each infected bovine, a volume of 0·3 mL of the infected blood was injected intraperitoneally (IP) into each of two OF1 mice. The injected mice were monitored for development of parasitaemia,

with each positive mouse considered as an isolate. Parasitaemic mice were euthanized and the blood collected for stabilate production. Six T. congolense strains were isolated from tsetse flies in the South Luangwa National Park in Zambia. The South Luangwa National Park is a protected game area where wildlife acts as reservoirs of the trypanosomes. Tsetse flies (Glossina morsitans morsitans and G. pallidipes) were trapped MycoClean Mycoplasma Removal Kit using epsilon traps (13), and live flies were dissected to determine their infection status. The mouthparts of tsetse flies, infected with trypanosomes in both

the midgut and mouthparts, were injected intraperitoneally (I.P.) into an immunosuppressed OF1 mouse (300 mg/kg Cyclophosphamide; Endoxan®, Baxter SA, Lessines, Belgium). The injected mice were then monitored for the development of parasitaemia, with each positive mouse considered as an isolate. Parasitaemic mice were euthanized and the blood collected for stabilate production. Finally, six T. congolense strains were isolated from buffalos belonging to herds that were selected randomly for tuberculosis testing in the Hluhluwe-iMmfolozi Park located in the KwaZulu-Natal Province of South Africa. From each of the 132 buffalo sampled, a volume of 0·3 mL of jugular blood was injected IP into each of two OF1 mice. The injected mice were then monitored as described previously, and stabilates were prepared from the blood of positive mice. The virulence of the T. congolense isolates, all belonging to the Savannah subgroup (14), was determined using a standard protocol in OF1 mice (9). All strains were at their fifth or sixth passages in mice.