First in vitro isolation of Besnoitia besnoiti from chronically infected cattle in Germany |
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Authors: | G Schares W Basso M Majzoub HCE Cortes A Rostaher J Selmair W Hermanns FJ Conraths NS Gollnick |
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Institution: | 1. Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Institute of Epidemiology, Wusterhausen, Germany;2. Laboratorio de Inmunoparasitología, Facultad de Ciencias Veterinarias, Universidad Nacional de La Plata, La Plata, Argentina;3. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina;4. Institut of Veterinary Pathology, Ludwig-Maximilians-Universität, Munich, Germany;5. Laboratório de Parasitologica, ICAM, Núcleo da Mitra, Universidade de Évora, Portugal;6. Medizinische Kleintierklinik, Ludwig-Maximilians-Universität, Munich, Germany;g Inning am Holz, Germany;h Clinic for Ruminants, Ludwig-Maximilians-Universität, Munich, Germany |
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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. |
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Keywords: | Besnoitia besnoiti Cell culture In vitro Ribosomal RNA gene Internal transcribed spacer 1 |
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