title: In vitro cultivation of a south African isolate of an Anaplasma sp. in tick cell cultures.
abstract: This paper describes the first successful in vitro cultivation of a South African isolate of an Anaplasma sp., initially thought to be Anaplasma marginale, in the continuous tick cell line IDE8. Blood from a bovine naturally infected with A. marginale kept on the farm Kaalplaas (28 degrees 08' E, 25 degrees 38' S) was collected, frozen, thawed and used as inoculum on confluent IDE8 cell cultures. Twenty days after culture initiation small intracellular colonies were detected in a Cytospin smear prepared from culture supernatant. Cultures were passaged on Day 34. Attempts to infect IRE/CTVM18 cell cultures with the Kaalplaas isolate derived from IDE8 cultures failed, whereas a reference stock of A. marginale from Israel infected IRE/CTVM18 tick cell cultures. Attempts to infect various mammalian cell lines (BA 886, SBE 189, Vero, L 929, MDBK) and bovine erythrocytes, kept under various atmospheric conditions, with tick cell-derived Anaplasma sp. or the Israeli strain of A. marginale failed. Molecular characterization revealed that the blood inoculum used to initiate the culture contained both A. marginale and Anaplasma sp. (Omatienne) whereas the organisms from established cultures were only Anaplasma sp. (Omatjenne).
authors.affiliation: Onderstepoort Veterinary Institute, Private Bag X5, Onderstepoort 0110, South Africa. ZweygarthE@arc.agric.za
authors.forename: E
authors.initials: E
authors.lastname: Zweygarth
authors.forename: A I
authors.initials: AI
authors.lastname: Josemans
authors.forename: A M
authors.initials: AM
authors.lastname: Spickett
authors.forename: H C
authors.initials: HC
authors.lastname: Steyn
authors.forename: J
authors.initials: J
authors.lastname: Putterill
authors.forename: P C
authors.initials: PC
authors.lastname: Troskie
authors.forename: M S
authors.initials: MS
authors.lastname: Mtshali
authors.forename: L
authors.initials: L
authors.lastname: Bell-Sakyi
authors.forename: V
authors.initials: V
authors.lastname: Shkap
authors.forename: L
authors.initials: L
authors.lastname: Fish
authors.forename: K M
authors.initials: KM
authors.lastname: Kocan
authors.forename: E F
authors.initials: EF
authors.lastname: Blouin
mesh_terms: D000709:Anaplasma / Q000145:classification / Q000254:growth & development* / Q000302:isolation & purification; D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D004269:DNA, Bacterial / Q000737:chemistry; D004912:Erythrocytes / Q000382:microbiology* / Q000648:ultrastructure; D018884:Ixodes / Q000166:cytology / Q000382:microbiology*; D008854:Microscopy, Electron / Q000662:veterinary; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary
chemical_list: D004269:DNA, Bacterial