In vitro cultivation of a newly recognized Babesia sp. in dogs in North Carolina

A novel large Babesia sp. from an infected dog was cultivated in vitro by microaerophilous stationary phase culture methodology. A primary culture initiated in enriched RPMI-1640 medium supplemented with 40% canine serum and incubated in a 2% oxygen environment supported parasite growth in vitro. Su...

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Bibliographic Details
Published in:Veterinary parasitology 2008-02, Vol.151 (2), p.150-157
Main Authors: Lehtinen, Lauren E., Birkenheuer, Adam J., Droleskey, Robert E., Holman, Patricia J.
Format: Article
Language:English
Subjects:
animal pathogens
Animals
Babesia
Babesia - classification
Babesia - genetics
Babesia - growth & development
Babesia - ultrastructure
Babesia sp
babesiosis
Babesiosis - parasitology
Babesiosis - veterinary
Base Sequence
blood serum
Canine babesiosis
carbon dioxide
Cells, Cultured
cryopreservation
culture media
dog diseases
Dog Diseases - parasitology
Dogs
Female
Genes, rRNA - genetics
HL-1 medium
in vitro studies
Microaerophilous stationary phase culture
microbial growth
Microscopy, Electron, Transmission - veterinary
Molecular Sequence Data
mortality
new species
North Carolina
nucleotide sequences
oxygen
parasitemia
ribosomal RNA
RPMI-1640
sequence analysis
Small subunit ribosomal RNA gene
Species Specificity
Transmission electron microscopy
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Summary:A novel large Babesia sp. from an infected dog was cultivated in vitro by microaerophilous stationary phase culture methodology. A primary culture initiated in enriched RPMI-1640 medium supplemented with 40% canine serum and incubated in a 2% oxygen environment supported parasite growth in vitro. Subsequent subcultures into enriched HL-1 medium with 20% fetal bovine serum also supported parasite propagation. Cultures were successfully introduced to 5% carbon dioxide in air atmosphere at passage 4. To date, the parasites have been continuously cultured through 35 passages, although the parasitemias are low, ranging from 0.2 to 0.3%. Parasites cultured in RPMI with canine serum were cryopreserved and successfully recovered from liquid nitrogen storage. The small subunit ribosomal rRNA gene sequence was identical in blood-derived and culture-derived parasites, differing in a single base position from the previously reported sequence for this Babesia sp. The ultrastructure of the parasite was consistent with that of other large Babesia spp., except that the spherical body contained numerous round particles unlike the inclusions previously described in Babesia spp.
ISSN:0304-4017
1873-2550
DOI:10.1016/j.vetpar.2007.10.022