Abstract:
The objective of our study was identification and molecular characterisation of piroplasms and
rickettsias occurring in brown (Parahyaena brunnea) and spotted hyaenas (Crocuta crocuta) from
various localities in Namibia and South Africa. Whole blood (n=59) and skin (n=3) specimens from
brown (n=15) and spotted hyaenas (n=47) were screened for the presence of Babesia, Theileria,
Ehrlichia and Anaplasma species using the Reverse Line Blot (RLB) hybridization technique. PCR
products of 52/62 (83.9%) of the specimens hybridized only with the Theileria/Babesia genus-specific
probes and not with any of the species-specific probes, suggesting the presence of a novel species or
variant of a species. No Ehrlichia and/or Anaplasma species DNA could be detected. Parasite 18S
rRNA gene of brown (n=3) and spotted hyaena (n=6) specimens was subsequently amplified, cloned
and the recombinants sequenced. Homologous sequence searches of databases indicated that the
obtained sequences were most closely related to B. lengau, originally described from cheetahs
(Acinonyx jubatus). Observed sequence similarities were subsequently confirmed by phylogenetic
analyses which showed that the obtained hyaena sequences formed a monophyletic group with B.
lengau, B. conradae and sequences previously isolated from humans and wildlife in the western USA.
Within the B. lengau clade, the obtained sequences and the published B. lengau sequences grouped
into six distinct groups, of which groups I to V represented novel B. lengau genotypes and/or gene
variants. We suggest that these genotypes cannot be classified as new Babesia species, but rather as
variants of B. lengau. This is the first report of occurrence of piroplasms in brown hyaenas.
