1. First in vitro isolation of Besnoitia besnoiti from chronically infected cattle in Germany
- Author
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Schares, G., Basso, W., Majzoub, M., Cortes, H.C.E., Rostaher, A., Selmair, J., Hermanns, W., Conraths, F.J., and Gollnick, N.S.
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BESNOITIOSIS , *APICOMPLEXA , *DIAGNOSTIC microbiology , *PATHOGENIC microorganisms , *PATHOGENIC protozoa , *MOLECULAR parasitology , *VETERINARY epidemiology , *VETERINARY protozoology , *NUCLEOTIDE sequence - Abstract
Abstract: Besnoitia besnoiti was in vitro isolated during the first recorded outbreak of bovine besnoitiosis in Germany. Molecular characterization of the new isolate, named Bb-GER1, revealed almost 100% identity with other B. besnoiti isolates obtained in Portugal, Spain, Israel or South Africa, when partial sequences of the 18S ribosomal RNA gene, of the internal transcribed spacer 1 and of the 5.8S RNA gene were compared. Cystozoites obtained from skin tissue of one bull were infectious for γ-interferon knockout (GKO) mice by intraperitoneal (ip) inoculation. Tachyzoites were detected in the peritoneal cavity, spleen, liver and lung of the mice 5 days post-infection. The parasite could be maintained in GKO mice by ip inoculation for at least 5 passages. Peritoneal washings containing tachyzoites were obtained from infected mice and used to infect five cell lines (Vero, MARC-145, NA42/13, BHK21, KH-R). The best growth of tachyzoites was observed in BHK21 cells, but replication occurred to a smaller extent also in MARC-145, NA42/13 and KH-R cells. Subsequent comparative analyses revealed that after direct infection of these cell lines with cystozoites derived from bovine skin, the growth was best in NA42/13 cells. Considerable replication was also observed in the BHK21 and KH-R cell lines. Our observations on the growth characteristics of Bb-GER1 partially contrast those for other isolates. The preferential growth in particular cell lines may be characteristic for particular B. besnoiti isolates. A potential association between growth properties and differences in virulence remains to be established. This is the first in vitro isolation of B. besnoiti from cattle in Germany. [Copyright &y& Elsevier]
- Published
- 2009
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