Characterisation of South African field Ehrlichia ruminantium using multilocus sequence typing.

Abstract

Heartwater, one of the major tick-borne diseases of some domestic and wild ruminants in Africa, is caused byEhrlichia ruminantium. The genetic diversity ofE. ruminantiumisolates renders the available vaccine ineffective against certain virulent isolates. To better understand theE. ruminantiumgenotypes in South Africa, a total of 1004Amblyomma hebraeumtick deoxyribonucleic acid (DNA) samples from cattle in three South African provinces were tested by pCS20 Sol1 real-time polymerase chain reaction (qPCR) and characterised by multilocus sequence typing (MLST) using five housekeeping genes. Out of 1004 samples tested, 222 (22%) were positive forE. ruminantium. The occurrence ofE. ruminantiumin Mpumalanga, KwaZulu-Natal and Limpopo provinces was 19%, 22% and 27%, respectively. TheE. ruminantiumpositive samples were screened for housekeeping genes and sequenced. Phylogenetic analysis revealed three main lineages: clade 1 made up of worldwide isolates (eastern, southern Africa, and Caribbean isolates), clade 2 comprised only West African isolates and clade 3 consisted of Omatjenne, Kümm2 and Riverside. Some study sample sequences were not identical to any of the reference isolates. However, they could all be grouped into the worldwide clade. Genetic variation in the sequenced regions was observed in the form of single nucleotide polymorphisms (SNPs). Using MLST to characteriseE. ruminantiumfield isolates allowed the South African genotypes to be clearly distinguished from the distinct West African isolates.Contribution:Characterisation ofE. ruminantiumfield isolates is important for the control of heartwater and contributes to preliminary knowledge required for the development of a more practical vaccine against heartwater.