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Background  Haemogregarines comprise a large group of apicomplexan blood parasites. In 1996 all anuran haemogregarines still in the genus  Haemogregarina  Danilewsky, 1885 were reassigned to the genus  Hepatozoon  Miller, 1908. Most (11/15, 73%) African anuran  Hepatozoon  species have been recorded from the family Bufonidae, however, all these are recorded from only two host species,  Amietophrynus mauritanicus  (Schlegel, 1841) and  Amietophrynus regularis  (Reuss, 1833) from Northern and central Africa. To the authors’ knowledge the only description of an anuran haemogregarine from South Africa is  Hepatozoon theileri  (Laveran, 1905), parasitising  Amietia quecketti  (Boulenger, 1895).    Methods  Thin blood smears for morphometrics and whole blood for molecular work, were collected from 32  Amietophrynus garmani  (Meek, 1897), 12  Amietophrynus gutturalis  (Power, 1927), and nine  Amietophrynus maculatus  (Hallowell, 1854), in Ndumo Game Reserve and Kwa Nyamazane Conservancy, KwaZulu-Natal, South Africa. Smears were Giemsa-stained, screened for haemogregarines, parasite stages measured, compared to each other and to other described African bufonid haemogregarines. Parasite 18S rDNA was amplified using two apicomplexan-specific primer sets, HepF300/HepR900 and 4558/2733. Resulting sequences of the haemogregarine isolates from the three  Amietophrynus  species were compared with each other and to comparative haemogregarine sequences selected from GenBank.    Results  Morphological characteristics of parasite stages, in particular characteristically capped mature gamont stages, and molecular findings, supported all three haemogregarine isolates from all three  Amietophrynus  species to be the same, a species of  Hepatozoon , and furthermore different morphologically from other previously recorded bufonid  Hepatozoon  species. The haemogregarine fell within a clade comprising other anuran  Hepatozoon  species and furthermore, within a monophyletic sub-clade along with  H. theileri  and are described as  Hepatozoon ixoxo  sp. nov.    Conclusions  This is the first morphological and molecular account of  Hepatozoon  species within the family Bufonidae from South Africa, a study hoped to encourage the redescription and molecular analysis of those  Hepatozoon  species described in the past from  Amietophrynus  species, as well as to promote the use of both morphological and molecular characteristics in  Hepatozoon  species descriptions. This will aid in comprehensive  Hepatozoon  descriptions, which along with the use of phylogenetic analysis will give a better indication of these parasites possible vectors and life cycle dynamics.

parasites and vectors

Hepatozoon species (Adeleorina: Hepatozoidae) of African bufonids, with morphological description and molecular diagnosis of Hepatozoon ixoxo sp. nov. parasitising three Amietophrynus species (Anura: Bufonidae)