Epidemiology and tick-borne haemoparasite diversity amongst transhumant Zebu cattle in Karamoja region, Uganda

Abstract

This study was conducted to understand the occurrence, diversity and epidemiology of tickborne diseases (TBDs) in a transhumant pastoral area of Karamoja Region, Uganda. We used participatory epidemiology (PE), involving focus group discussions (n = 24) with livestock keepers, 30 key informant interviews, review of previous surveillance data, clinical examinations, and laboratory confirmation of cases of TBDs, to define and prioritise cattle diseases, evaluate current control activities, and identify constraints to the control of TBDs. The livestock keepers regarded TBDs, particularly East Coast fever (ECF) and anaplasmosis, as the most important health problems in their cattle, based on morbidity and mortality rates, rates of transmission, treatment costs, difficulty in accessing the correct treatment, difficulty to control, and inadequate knowledge to manage the diseases. The main constraints to the control of TBDs were inadequate knowledge, inadequate veterinary services and limited availability of drugs. Hand picking of ticks was done by all pastoralists while hand spraying was done with under-strength acaricides, often at irregular intervals and with little acaricide wash. We determined the endemic status of TBDs in 20 randomly-selected cattle herds by estimating the proportion of annual ECF and anaplasmosis cases in different age groups of cattle using participatory approaches and clinical examinations, determining the diversity and abundance of ticks (161 cattle in 20 herds), and establishing the seroprevalence of antibodies to Theileria parva and Anaplasma marginale among cattle (n = 397 in 20 herds). Clinical examinations and informant interviews showed that TBDs affected all age groups of cattle. Two-thirds of the cattle were infested with moderate (37.3%, 11-50 ticks) to abundant (28.6%, > 50 ticks) numbers of ticks. Out of the 10,923 ticks collected, Rhipicephalus appendiculatus (54.4%) was the most abundant species followed by R. decoloratus (17.7%), Amblyomma variegatum (12%) and A. lepidum (11.6%). We collected tick species that are either rare in Uganda (A. lepidum, Hyalomma truncatum, A. gemma) or were not known to be present in the country (R. pulchellus). The true seroprevalence of antibodies to A. marginale was high (95.1%, 95% confidence interval [CI] 91.3% - 98.5%), while that of T. parva was low (16.5%, 95% CI 12.9% - 19.6%). We determined the presence of tick-borne haemoparasites among cattle and the prevalence of infections using reverse line blot (RLB) hybridisation and quantitative real-time polymerase chain reaction (qPCR), and characterised Theileria and Babesia species by sequence and phylogenetic analyses of the full-length 18S rRNA gene. The RLB hybridisation assay demonstrated the presence of tick-borne haemoparasites in most cattle (99.6%), mostly as mixed infections (97.5%). The most frequently-detected species were Theileria mutans (88.3%, 95% CI: 84.6-92.1%), A. marginale (73.8%: 68.3-79.2%), T. velifera (71.3%: 65.8-76.7%) and Anaplasma sp. Omatjenne (63.3%: 57.5-68.8%). Other virulent pathogens, namely Babesia bigemina (5.0%) and T. parva (2.9%), were also detected with RLB, but Ehrlichia ruminantium was not detected. The proportions of qPCR positive samples were 82.9% (A. marginale), 12.1% (A. centrale), 3.3% (T. parva), and 1.7% (E. ruminantium). Variations (5 to 9 nucleotides) in the 18S rRNA gene sequences of B. bigemina were identified as compared with previously published sequences. We assessed the 16S rRNA gene phylogeny of Anaplasma species from cattle and analysed the msp1α gene sequences of A. marginale to identify genotypes. There was genetic heterogeneity within A. marginale in cattle in the study area. Most A. marginale sequences (16/19) were closely related (99- 100% identities) and clustered with A. marginale strain Veld from cattle in South Africa, with strong bootstrap support, while three sequences clustered (100% identity) with A. marginale strain from Virginia, USA. The A. centrale sequences were closely related (100% identity) and clustered with the Israel vaccine strain. We found four different kinds of MSP1a tandem repeat sequences (UP39-F-M2-3) that correspond to one A. marginale strain unique to Uganda. One tandem repeat (UP39) was unique to Karamoja cattle. The findings from this study provide knowledge on the diversity and epidemiology of TBDs, which can be used to support diagnosis and strategic control of TBDs, and consequently improve cattle productivity and the livelihoods of pastoralists. More effective control and prevention measures against ticks and TBDs should urgently be implemented in the region. There is need to facilitate and promote immunisation of cattle against ECF. Given the scarcity of veterinarians in the region, the activities of community-based animal health workers [CAHWs] should be strengthened. The local governments in Karamoja Region should facilitate a more favourable business environment to attract private companies, who could scale-up veterinary drug supplies to the region. Pastoralists should be sensitised to improve their ecological knowledge of the seasonal activity of ticks for strategic tick control.