Tuesday, 16 November 2004

Prevalence and in vitro isolation of Anaplasma sp. in black-legged ticks collected from white-tailed deer

Timothy J. Kurtti, kurtt001@umn.edu1, Jariyanart K. Gaywee, jariyanart@yahoo.com2, Meghan J. Lynch1, and Ulrike G. Munderloh, munde001@umn.edu1. (1) University of Minnesota, Entomology, 219 Hodson Hall, 1980 Folwell Av, Saint Paul, MN, (2) Armed Forces Research Institute of Medical Sciences, Microbiology, 315/6 Rajvithee Road, Rajthawee, Bangkok, Thailand

White tailed deer are important hosts for black-legged ticks in the north central US and are natural reservoirs for Anaplasma and Ehrlichia species. The goals of this study were to investigate the endemicity of granulocytic Anaplasma in ticks in an area with high deer density, to obtain culture isolates directly from ticks, and to determine the relative abundance of the presumed non-human infectious variant of Anaplasma phagocytophilum. We used tick cell cultures to isolate Anaplasma from ticks collected from deer hunted at Camp Ripley, Minnesota. PCR with broad range primers indicated that 50% (n=22) of ticks were positive for Anaplasma or Ehrlichia. Within one week after removal from deer, 30 partially engorged ticks were surface disinfected and their organs extirpated for cultivation in multi-well plates seeded with Ixodes scapularis cell line ISE6. Seven cultures became positive for Anaplasma and resulted in 5 continuous Anaplasma isolates maintained in ISE6 cells. DNA sequencing of a portion of the 16S rRNA gene identified all culture isolates as the AP-Variant 1 of A. phagocytophilum.

Species 1: Acari Ixodidae Ixodes scapularis (black-legged tick)
Species 2: Rickettsiales Anaplasmataceae Anaplasma phagocytophilum (HGA agent)
Keywords: tick, culture

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