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. 2023 Oct 4;10(10):605.
doi: 10.3390/vetsci10100605.

Molecular Detection of Rickettsia hoogstraalii in Hyalomma anatolicum and Haemaphysalis sulcata: Updated Knowledge on the Epidemiology of Tick-Borne Rickettsia hoogstraalii

Aneela Aneela  1 , Mashal M Almutairi  2 , Abdulaziz Alouffi  3 , Haroon Ahmed  4 , Tetsuya Tanaka  5 , Itabajara da Silva Vaz Junior  6 , Shun-Chung Chang  7 , Chien-Chin Chen  8   9   10   11 , Abid Ali  1
Affiliations

Molecular Detection of Rickettsia hoogstraalii in Hyalomma anatolicum and Haemaphysalis sulcata: Updated Knowledge on the Epidemiology of Tick-Borne Rickettsia hoogstraalii

Aneela Aneela et al. Vet Sci. .

Abstract

Ticks are hematophagous ectoparasites that transmit pathogens to animals and humans. Updated knowledge regarding the global epidemiology of tick-borne Rickettsia hoogstraalii is dispersed, and its molecular detection and genetic characterization are missing in Pakistan. The current study objectives were to molecularly detect and genetically characterize Rickettsia species, especially R. hoogstraalii, in hard ticks infesting livestock in Pakistan, and to provide updated knowledge regarding their global epidemiology. Ticks were collected from livestock, including goats, sheep, and cattle, in six districts of Khyber Pakhtunkhwa (KP) Pakistan. Overall, 183 hosts were examined, of which 134 (73.2%), including goats (number = 39/54, 72.2%), sheep (23/40, 57.5%), and cattle (71/89, 80%) were infested by 823 ticks. The most prevalent tick species was Rhipicephalus microplus (number = 283, 34.3%), followed by Hyalomma anatolicum (223, 27.0%), Rhipicephalus turanicus (122, 14.8%), Haemaphysalis sulcata (104, 12.6%), Haemaphysalis montgomeryi (66, 8.0%), and Haemaphysalis bispinosa (25, 3.03%). A subset of 210 ticks was selected and screened for Rickettsia spp. using PCR-based amplification and subsequent sequencing of rickettsial gltA and ompB fragments. The overall occurrence rate of R. hoogstraalii was 4.3% (number = 9/210). The DNA of Rickettsia was detected in Hy. anatolicum (3/35, 8.5%) and Ha. sulcata (6/49, 12.2%). However, no rickettsial DNA was detected in Rh. microplus (35), Rh. turanicus (35), Ha. montgomeryi (42), and Ha. bispinosa (14). The gltA and ompB fragments showed 99-100% identity with R. hoogstraalii and clustered phylogenetically with the corresponding species from Pakistan, Italy, Georgia, and China. R. hoogstraalii was genetically characterized for the first time in Pakistan and Hy. anatolicum globally. Further studies should be encouraged to determine the role of ticks in the maintenance and transmission of R. hoogstraalii in different hosts.

Keywords: Ixodidae; Pakistan; Rickettsia hoogstraalii; ticks.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Map showing the locations (black triangles) of tick collection in specific districts of Khyber Pakhtunkhwa (KP), Pakistan.
Figure 2
Figure 2
A maximum likelihood phylogenetic tree of R. hoogstraalii was constructed based on the gltA fragment. R. canadensis was used as an outgroup. The bootstrap values (1000-replication) are shown at each node. The obtained sequence (OR392758) of the present study is marked in bold and underlined font.
Figure 3
Figure 3
A maximum likelihood phylogenetic tree of R. hoogstraalii was constructed based on the ompB partial fragment. R. prowazekii was used as the outgroup. Bootstrap values (1000-replication) are shown at each node. The obtained sequence (OR392759) of the present study is marked in bold and underlined font.
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