Fig 1.
Ticks were collected from cattle in a farm in Xi Tang Town, Yueyang City, Hunan Province in Southern China from August 5 to August 22, 2022. The map was constructed using ArcGIS 10.6 software. The basemap shapefiles were downloaded from national platform for common geospatial information services (tianditu.gov.cn).
Fig 2.
The oviposition of a female Rhipicephalus microplus.
(A) Photographs taken on the 3rd and 7th day of oviposition, respectively. (B) Micrographs of tick eggs (top) and laboratory hatched larvae (bottom).
Table 1.
PCR primers used to amplify Rickettsia, Anaplasma and Ehrlichia species.
Fig 3.
Photos of the dorsal (left) and ventral (right) sides of Rhipicephalus microplus under a dissecting microscope.
(A) An adult female tick. (B) An adult male tick.
Fig 4.
Sample pooling information.
Fig 5.
Phylogenetic tree of Rhipicephalus microplus ticks.
The tree was constructed using the partial 16S rRNA gene (229 bp) of ticks based on the Maximum Likelihood (ML) method with the Kimura 2-parameter model in MEGA 7.0. Rhipicephalus sanguineus sequence was used for outgroup in the tree. The R. microplus sequences obtained in this study were marked with dots and have been submitted to the GenBank with accession numbers: OQ975295–OQ975297.
Table 2.
Prevalence of Rickettsia, Anaplasma, and Ehrlichia in Rhipicephalus microplus ticks collected from cattle in Hunan Province, China from August 5 to August 22, 2022.
Table 3.
The rickettsial sequences in the GenBank sharing the highest homology with the rickettsial sequences obtained in this study.
Fig 6.
Phylogenetic tree of Rickettsia based on the concatenated sequences of htrA (365 bp), rrs (1241 bp), ompA (551 bp), gltA (604 bp), and ompB (344 bp) genes.
Phylogenetic tree was constructed based on the Maximum Likelihood (ML) method with the Kimura 2-parameter model in MEGA 7.0. Bootstrap values (inferred from 1,000 replicates) >60% were indicated. The Rickettsia sequences obtained in this study were marked with dots. For the Rickettsia species without complete genome sequences, the GenBank accession nos. in the order of htrA, rrs, ompA, gltA and ompB are KY617773, KY617772, KU853021, KU853023, and KY617776 for C. Rickettsia xinyangensis; MG906673, MT747412, MN026548, MN026549, and MT747415 for C. Rickettsia longicornii; KT187396, MT062904, KT326194, KT187394, and JF758826 for Rickettsia vini.
Fig 7.
Phylogenetic trees of Anaplasma based on the partial rrs (470 bp) gene and groEL (205 bp) genes.
Phylogenetic trees were constructed based on the Maximum Likelihood (ML) method with the Kimura 2-parameter model in MEGA 7.0. Bootstrap values (inferred from 1,000 replicates) more than 60% were indicated. Wolbachia pipientis sequences were used for outgroup in the trees. The Anaplasma sequences obtained in this study were marked with dots.
Fig 8.
Phylogenetic trees of Ehrlichia with the partial rrs (430 bp) and groEL (461 bp) genes.
Phylogenetic trees were constructed using the Maximum Likelihood (ML) method with the Kimura 2-parameter model in MEGA 7.0. Bootstrap values (inferred from 1,000 replicates) more than 60% were indicated. Anaplasma marginale sequences were used for outgroup in the trees. The Ehrlichia sequences obtained in this study were marked with dots.