Wide distribution of Mediterranean and African Spotted fever agents and the first identification of Israeli Spotted Fever agent in ticks in Uganda

Rickettsia microorganisms are causative agents of several neglected emerging infectious diseases transmitted to humans by ticks among other arthropod vectors. In this study, ticks were collected from four geographical regions of Uganda, pooled in sizes of 1-179 ticks based on location, tick species, life stage, host, and time of collection, and were tested by real time PCR for Rickettsia species harboured. The tick pools were tested with primers targeting gltA, 17kDa and ompA genes, followed by Sanger sequencing of 17kDa and ompA genes. Of the 471 tick pools tested, 116 (24.6%) were positive for Rickettsia spp. by the gltA primers. The prevalence of Rickettsia varied by district with Gulu recording the highest (30.1%) followed by Luwero (28.1%) and Kasese had the lowest (14%). Tick pools with highest positivity rates were from livestock (cattle, goats, sheep, and pigs), 26.9%, followed by vegetation 23.1% and pets (dogs and cats) 19.7%. Of 116 gltA-positive tick pools, 86 pools were positive using 17kDa primers of which 48 purified PCR products were successfully sequenced. The predominant Rickettsia spp. identified was R. africae (n=15) in four tick species, followed by R. conorii (n=5) in three tick species (Haemaphysalis elliptica, Rhipicephalus appendiculatus, and Rh. decoloratus). Rickettsia conorii subsp. israelensis was detected in one tick pool. These findings indicate that multiple Rickettsia spp. capable of causing human illness are circulating in the four diverse geographical regions of Uganda including new strains previously known to occur in the Mediterranean region. Physicians should be informed about Rickettsia spp. infections as potential causes for acute febrile illnesses in these regions. Continued and expanded surveillance is essential to further identify and locate potential hotspots with Rickettsia spp. of concern. Author SummaryTick-borne rickettsioses are emerging infectious diseases of public health importance worldwide. Spotted fever rickettsioses transmitted by ticks can cause mild to severe human illness depending on the Rickettsia spp. and co-morbidities. Their diagnosis is challenging due to non-specific symptoms particularly in limited resource settings. Little is known about their prevalence in Uganda. Using entomological and molecular tools, we surveyed and studied tick-borne spotted fever rickettsioses in five districts from four diverse eco-regions of Uganda. Overall, 24.6% (116/471) tick pools were positive for Rickettsia species. By sequencing the 17kDa and ompA genes of Rickettsia, we identified R. africae as the most common agent, followed by R. conorii and R. conorii subsp. israelensis. The findings indicate multiple Rickettsia spp. that can cause febrile illness in humans are circulating in the four geographically diverse regions of Uganda. Physicians should be aware these agents are potential causes of febrile illness in these areas, particularly in individuals who encounter livestock or their grazing areas.