In Uganda, the role of ticks in zoonotic disease transmission is not well described, partly, due to limited available information on tick diversity. This study aimed to identify the tick species that infest cattle. Between September and November 2017, ticks (
n
= 4362) were collected from 5 districts across Uganda (Kasese, Hoima, Gulu, Soroti, and Moroto) and identified morphologically at Uganda Virus Research Institute. Morphological and genetic validation was performed in Germany on representative identified specimens and on all unidentified ticks. Ticks were belonging to 15 species: 8
Rhipicephalus
species (
Rhipicephalus appendiculatus
,
Rhipicephalus evertsi evertsi
,
Rhipicephalus microplus
,
Rhipicephalus decoloratus
,
Rhipicephalus afranicus
,
Rhipicephalus pulchellus
,
Rhipicephalus simus
, and
Rhipicephalus sanguineus
tropical lineage); 5
Amblyomma
species (
Amblyomma lepidum
,
Amblyomma variegatum
,
Amblyomma cohaerens
,
Amblyomma gemma
, and
Amblyomma paulopunctatum
); and 2
Hyalomma
species (
Hyalomma rufipes
and
Hyalomma truncatum
). The most common species were
R. appendiculatus
(51.8%),
A. lepidum
(21.0%),
A. variegatum
(14.3%),
R. evertsi evertsi
(8.2%), and
R. decoloratus
(2.4%)
. R. afranicus
is a new species recently described in South Africa and we report its presence in Uganda for the first time. The sequences of
R. afranicus
were 2.4% divergent from those obtained in Southern Africa. We confirm the presence of the invasive
R. microplus
in two districts (Soroti and Gulu). Species diversity was highest in Moroto district (
p
= 0.004) and geographical predominance by specific ticks was observed (
p
= 0.001). The study expands the knowledge on tick fauna in Uganda and demonstrates that multiple tick species with potential to transmit several tick-borne diseases including zoonotic pathogens are infesting cattle.