Molecular biological comparison of different Besnoitia species and stages from different countries.

Besnoitia besnoiti tissue cysts from a recent outbreak in cattle in Germany were characterized with respect to their internal transcribed spacer regions 1, 2, and 18S rDNA gene sequences. These results were compared with own sequences of an Israelian isolate of B. besnoiti and of Besnoitia jellisoni cystozoites stored for years in liquid nitrogen. Furthermore, material was studied that was obtained from white mice (Balb/C) that had been successfully infected by intraperitoneal infection of fresh cystozoites from the German outbreak. All results were then compared and discussed with respect to databank sequences of other Besnoitia species. Comprehensive phylogenetic studies of B. besnoiti isolates from Germany revealed almost identical sequence alignments when compared to previously sequenced B. besnoiti isolates from Israel and Spain. More importantly, phylogenetic analysis revealed two distant clusters of Besnoitia species: the first one includes Besnoitia akodoni, Besnoitia darlingi, and Besnoitia oryctofelisi, while the second cluster includes B. besnoiti, Besnoitia bennetti, Besnoitia tarandi, and the Besnoitia species of rodents (B. jellisoni). The also B. jellisoni named species of the GenBank (AF 076860) must be another one, since our strain derives directly from Frenkel. These findings give strong hints that B. besnoiti has a cycle between rodents and a predator and that cattle and other are only accidental hosts.

Another African disease in Central Europa: Besnoitiosis of cattle. I. Light and electron microscopical study.

The paper reports the first detection of besnoitiosis of cattle in Germany. Just 2 years after the first appearance of the African Bluetongue disease (BTD) of cattle in Central Europe, another African agent of disease has arrived in Germany. While it was proven that the BTD virus was transmitted (after its first appearance) by endemic midges of the genus Culicoides (C. obsoletus, C. pulicaris), nothing is known, how the infectious stages of Besnoitia besnoiti-a member of the so-called cyst-forming coccidia-found their way to a herd in Southern Germany. The infected animals showed all characteristic clinical symptoms of besnoitiosis such as hyposclerodermia, hyperkeratosis, alopecia, and whitish tissue cysts in subcutaneous tissues as well as in the cornea. These cysts had diameters of up to 3 mm and consisted of a dense outer layer (=secondary cyst wall), which surrounded a host cell, that had been enormously enlarged by an inner parasitophorous vacuole containing thousands of 7-9 x 2 mum sized, banana-shaped cyst merozoites (=cystozoites, bradyzoites).Their fine structure was identical to that of published stages of B. besnoiti. During cyst development, the nucleus of the host cell had been hypertrophied and had apparently undergone several divisions, since many flattened, but very large nuclei were seen in light and electron microscopy. Thus, this study proves the arrival of another serious agent of disease of ruminants in Central Europe-a fact which is especially important, since in this species, there is neither information on the way of transmission from animal to animal nor exists concrete information on an efficacious therapy or on the modalities of its import into Germany.