The Rhipicephalus appendiculatus tick vector of Theileria parva is absent from cape buffalo (Syncerus caffer) populations and associated ecosystems in northern Uganda.

Rhipicephalus appendiculatus is the major tick vector of Theileria parva, an apicomplexan protozoan parasite that causes the most economically important and lethal disease of cattle in East and central Africa. The African cape buffalo (Syncerus caffer) is the major wildlife host of T. parva from southern Uganda and Kenya to southern Africa. We show herein that R. appendiculatus appears to be absent from the two largest national parks in northern Uganda. Syncerus caffer is common in both of these national parks, specifically Murchison falls (MFNP) and Kidepo Valley (KVNP). We re-confirmed the previously reported absence of T. parva in buffalo sampled in the two northern parks based on RLB data using a nested PCR based on the T. parva p104 gene. By contrast, T. parva-infected R. appendiculatus ticks and parasite-infected buffalo were present in Lake Mburo (LMNP) in South central Uganda. This suggests that the distribution of R. appendiculatus, which is predicted to include the higher rainfall regions of northern Uganda, may be limited by additional, as yet unknown factors.

Evaluation of non-centrifuged dried plasma spots versus centrifuged and non-centrifuged plasma for determination of HIV-1 viral load.

Accurate viral load measurement in plasma specimens is subject to the transport conditions applied since the stability of HIV-1 RNA can be at risk. Also, except during the primary infection, HIV is unlikely to be free in circulation because most patients produce specific antibodies in the weeks following primary infection. This study evaluated non centrifuged dried plasma spots versus centrifuged and non centrifuged plasma in the determination of HIV-1 viral load. A total of 40 patients infected with HIV were bled and three groups of samples were prepared from each patient. The first group was centrifuged at 1500×g for 20min, the second was not centrifuged but left to sediment by gravity for up to 3h, and the third was for dried plasma spots prepared from the same non centrifuged plasma. HIV RNA quantitation in plasma and dried plasma spots was evaluated by the Pearson correlation and a T-test. The three groups yielded average viral loads of 58,249; 83,355 and 116,963 copies/ml for centrifuged, non centrifuged and dried plasma spot samples respectively. The correlation for centrifuged versus non centrifuged was R(2)=0.78, that of centrifuged and dried plasma spots was R(2)=0.72 and finally R(2)=0.81 between non centrifuged and dried plasma spot samples. A significant difference in viral load results of centrifuged and DPS samples prepared from non centrifuged plasma was observed.

In Vitro Trypanocidal Activity of Antibodies to Bacterially Expressed Trypanosoma brucei Tubulin.

BACKGROUND: There are only four drugs for treating African trypanosomiasis, a devastating disease in sub-Saharan Africa. With slow discovery of better drugs, vaccination is viewed as the best method of control. We previously showed that antibodies to native Trypanosoma brucei brucei tubulin inhibit the growth of trypanosomes in culture. Here, we aimed to determine the effect of antibodies to bacterially expressed trypanosome tubulin on T. brucei brucei growth. METHODS: T. brucei brucei alpha and beta tubulin genes were individually expressed in Escherichia coli under the tryptophan promoter. Monoclonal tubulin antibodies reacted specifically with the expressed tubulins with no cross-reaction with the opposite tubulin. Rabbits were immunized with 450µg each of the concentrated recombinant tubulin, and production of antibodies assessed by ELISA and Western blotting. The effect of polyclonal antibodies on trypanosome growth was determined by culturing bloodstream T. brucei brucei in up to 25% of antisera. RESULTS: Low antisera dilutions (25%) from the immunized rabbits inhibited trypanosome growth. The most cytotoxic antisera were from one rabbit immunized with a mixture of both alpha and beta tubulins. However, the result was not reproduced in other rabbits and there was no apparent effect on growth at higher antisera dilutions. CONCLUSION: Antibodies to bacterially expressed trypanosome tubulin are not effective at killing cultured bloodstream trypanosomes.

Cryptosporidium sp. and Giardia sp. infections in mountain gorillas (Gorilla gorilla beringei) of the Bwindi Impenetrable National Park, Uganda.

For conservation purposes and because of growing ecotourism, some mountain gorilla (Gorilla gorilla beringei) populations have been habituated to humans. Fecal specimens (n = 100) of nonhabituated and human-habituated gorillas (5 populations; 6 age classes) were tested for Cryptosporidium sp. oocysts and Giardia sp. cysts by conventional staining and immunofluorescent antibody (IFA). Cryptosporidium sp. infections (prevalence 11%) were not restricted to very young gorillas but were observed in 3-yr-old to >12-yr-old gorillas; most of the infections (73%) occurred in human-habituated gorillas. The prevalence of Giardia sp. infections was 2%; 1 nonhabituated gorilla was concomitantly infected. Oocysts of Cryptosporidium sp. in the gorilla stools were morphologically, morphometrically, and immunologically undistinguishable from a bovine isolate of Cryptosporidium parvum oocysts. Mean concentration of Cryptosporidium sp. oocysts and Giardia sp. cysts in gorilla stools was 9.39x10(4)/g, and 2.49x10(4)/g, respectively. There was no apparent relationship between oocyst concentration and gorilla age, sex, or habituation status. Most Cryptosporidium sp. infections found in gorillas with closest proximity to people may be a result of the habituation process and ecotourism. This study constitutes the first report of Cryptosporidium sp. infections in the family Pongidae, in the free-ranging great apes, and in the species of gorilla.

Anaplasmosis in Uganda. II. Prevalence of bovine anaplasmosis.

The prevalence of bovine anaplasmosis was studied in 320 Zebu cattle randomly selected from three regions of Uganda (central, south-western and north-western) using DOT-ELISA, Western immunoblotting, Rapid Card Agglutination Test (RCAT), Capillary Tube Agglutination Test (CAT), Complement Fixation Test (CFT), and parasitological techniques. Dried blood on Whatman filter paper no. 1 was eluated in PBS 0.05% Tween 20 prior to testing at an initial dilution of 1:25. The incidence of parasitaemia ranged from 25% in the central region to 35% in the north-western region and the serological prevalence was lower in the central region and highest in the north-west. Prevalence rates assayed by DOT-ELISA and Western immunoblotting were 1.5-fold greater than those tested with RCAT and 3-fold greater than in CAT. The overall prevalence rates by DOT-ELISA and Western immunoblotting compared favourably with CFT data. The present data utilizing dried blood on filter papers indicate that there is a high prevalence of anaplasmosis in those regions of Uganda surveyed and it confirms our observations and those of others that collecting blood on filter papers is a suitable technique for large-scale screening and for seroepidemiological studies.