title: Comparative strain analysis of Anaplasma phagocytophilum infection and clinical outcomes in a canine model of granulocytic anaplasmosis.
abstract: A pilot study was conducted to determine whether existing human or canine strains of Anaplasma phagocytophilum would reproduce clinical disease in experimentally inoculated dogs similar to dogs with naturally acquired granulocytic anaplasmosis. Six hounds were inoculated intravenously with one human and two canine strains of A. phagocytophilum that were propagated in vitro in HL-60 cells or in infected autologous neutrophils. Infected dogs were monitored for lethargy, anorexia, petechiae, lymphadenopathy, and fever. Dogs were assessed for complete blood count (CBC), serum chemistry, and serology (IFA and SNAP® 4Dx®); for A. phagocytophilum blood load by quantitative polymerase chain reaction; and for cytokine production. Prominent clinical signs were generalized lymphadenopathy and scleral injection; only one dog developed fever lasting 4 days. Notable laboratory alterations included sustained leukopenia and thrombocytopenia in all dogs. A. phagocytophilum morulae were noted in blood between days 10 and 11, although all dogs retained A. phagocytophilum DNA in blood through day 60. All dogs seroconverted by days 10-15 by IFA, and by days 17-30 by SNAP 4Dx; cytokine analyses revealed 10-fold increases in interleukin-2 and interleukin-18 in the neutrophil-propagated 98E4 strain-infected dog. All A. phagocytophilum strains produced infection, although canine 98E4 strain reproduced clinical signs, hematologic changes, and inflammatory cytokine elevations most consistent with granulocytic anaplasmosis when recognized clinically. Therefore, this strain should be considered for use in future studies of A. phagocytophilum canine infection models.
authors.affiliation: Department of Molecular Pathobiology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA. dscorpio@jhmi.edu
authors.forename: Diana G
authors.initials: DG
authors.lastname: Scorpio
authors.forename: J Stephen
authors.initials: JS
authors.lastname: Dumler
authors.forename: Nicole C
authors.initials: NC
authors.lastname: Barat
authors.forename: Judith A
authors.initials: JA
authors.lastname: Cook
authors.forename: Christopher E
authors.initials: CE
authors.lastname: Barat
authors.forename: Brett A
authors.initials: BA
authors.lastname: Stillman
authors.forename: Kristen C
authors.initials: KC
authors.lastname: DeBisceglie
authors.forename: Melissa J
authors.initials: MJ
authors.lastname: Beall
authors.forename: Ramaswamy
authors.initials: R
authors.lastname: Chandrashekar
mesh_terms: D041081:Anaplasma phagocytophilum / Q000235:genetics / Q000276:immunology* / Q000472:pathogenicity; D000712:Anaplasmosis / Q000276:immunology / Q000382:microbiology*; D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood*; D058491:Bacterial Load; D002474:Cell-Free System; D016207:Cytokines / Q000097:blood; D004269:DNA, Bacterial / Q000097:blood; D004195:Disease Models, Animal; D004285:Dogs; D006098:Granulocytes / Q000382:microbiology*; D018922:HL-60 Cells; D006801:Humans; D007275:Injections, Intravenous; D008206:Lymphatic Diseases / Q000382:microbiology; D009504:Neutrophils / Q000382:microbiology; D010865:Pilot Projects; D060888:Real-Time Polymerase Chain Reaction; D013047:Specific Pathogen-Free Organisms
chemical_list: D000907:Antibodies, Bacterial; D016207:Cytokines; D004269:DNA, Bacterial