title: Sheep experimentally infected with a human isolate of Anaplasma phagocytophilum serve as a host for infection of Ixodes scapularis ticks.
abstract: Anaplasma phagocytophilum, first identified as a pathogen of ruminants in Europe, has more recently been recognized as an emerging tick-borne pathogen of humans in the U.S. and Europe. A. phagocytophilum is transmitted by Ixodes spp., but the tick developmental cycle and pathogen/vector interactions have not been fully described. In this research, we report on the experimental infection of sheep with the human NY-18 isolate of A. phagocytophilum which then served as a host for infection of I. scapularis nymphs and adults. A. phagocytophilum was propagated in the human promyelocytic cell line, HL-60, and the infected cell cultures were then used to infect sheep by intravenous inoculation. Infections in sheep were confirmed by PCR and an Anaplasma-competitive ELISA. Clinical signs were not apparent in any of the infected sheep, and only limited hematologic and mild serum biochemical abnormalities were identified. While A. phagocytophilum morulae were rarely seen in neutrophils, blood film evaluation revealed prominent large granular lymphocytes, occasional plasma cells, and rare macrophages. Upon necropsy, gross lesions were restricted to the lymphoid system. Mild splenomegaly and lymphadenomegaly with microscopic evidence of lymphoid hyperplasia was observed in all infected sheep. Female I. scapularis that were allowed to feed and acquire infection on each of the 3 experimentally infected sheep became infected with A. phagocytophilum as determined by PCR of guts (80-87%) and salivary glands (67-100%). Female I. scapularis that acquired infection as nymphs on an experimentally infected sheep transmitted A. phagocytophilum to a susceptible sheep, thus confirming transstadial transmission. Sheep proved to be a good host for the production of I. scapularis infected with this human isolate of A. phagocytophilum, which can be used as a model for future studies of the tick/pathogen interface.
authors.affiliation: Dept. of Veterinary Pathobiology, 250 McElroy Hall, Center for Veterinary Health Sciences, Oklahoma State University, Stillwater, OK 74078, USA. Katherine.Kocan@okstate.edu
authors.forename: Katherine M
authors.initials: KM
authors.lastname: Kocan
authors.forename: Ann T
authors.initials: AT
authors.lastname: Busby
authors.forename: Robin W
authors.initials: RW
authors.lastname: Allison
authors.forename: Melanie A
authors.initials: MA
authors.lastname: Breshears
authors.forename: Lisa
authors.initials: L
authors.lastname: Coburn
authors.forename: Ruth C
authors.initials: RC
authors.lastname: Galindo
authors.forename: Nieves
authors.initials: N
authors.lastname: Ayllón
authors.forename: Edmour F
authors.initials: EF
authors.lastname: Blouin
authors.forename: José
authors.initials: J
authors.lastname: de la Fuente
mesh_terms: D041081:Anaplasma phagocytophilum / Q000235:genetics / Q000276:immunology / Q000502:physiology*; D000818:Animals; D000942:Antigens, Bacterial / Q000276:immunology; D001096:Arachnid Vectors / Q000382:microbiology* / Q000821:virology; D002460:Cell Line; D004269:DNA, Bacterial / Q000235:genetics; D016873:Ehrlichiosis / Q000150:complications / Q000382:microbiology*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D041981:Gastrointestinal Tract / Q000382:microbiology; D006801:Humans; D018884:Ixodes / Q000382:microbiology* / Q000821:virology; D008297:Male; D023421:Models, Animal; D009504:Neutrophils / Q000382:microbiology; D009758:Nymph; D016133:Polymerase Chain Reaction; D012469:Salivary Glands / Q000382:microbiology; D012756:Sheep; D013984:Tick Infestations / Q000150:complications / Q000469:parasitology*
chemical_list: D000942:Antigens, Bacterial; D004269:DNA, Bacterial