Propagation of the Agent of HGE. Blood (EDTA anticoagulated) was obtained from a Nantucket patient with HGE whose infection was confirmed by blood smear and sequencing of a 16S rDNA amplification product (8), and 0.5 ml was inoculated i.p. into a splenectomized CD-1 mouse. Buffy coat preparations (equivalent to 1 ml of whole blood, centrifuged at 2500 x g for 10 min) were inoculated i.p. into two intact C3H/HeJ mice. Plasma (0.5 ml) was inoculated into one C3H/HeJ mouse as well. To determine whether ehrlichiae were present in the peripheral blood, thin blood smears were taken from the tails of rodents, dried, fixed for 2 min in absolute methanol, and stained with 1 ml Giemsa stock (Harleco Original Azure Blend; EM Diagnostic Systems, Princeton, NJ) diluted in 40 ml distilled water with 150 ptl of 0.5% Na2CO3 and 1.25 ml methanol for 1 hr or more (9

Ehrlichia chaffeensis Anderson, Dawson & Wilson, causative agent of human (predominantly monocytic) ehrlichiosis, was successfully transmitted experimentally by Amblyomma americanum (L.) to white-tailed deer, Odocoileus virginianus (Zimmerman). Deer were needle-exposed intravenously to E. chaffeensis in tissue-culture canine macrophage (DH82) cells, and 11 d later were exposed to laboratory-reared A. americanum larvae, nymphs, and adults for acquisition feeding. Three months after this feeding, naive deer and dogs were exposed to recently molted nymphs and adults. Attempted reisolation of the pathogen by way of tissue culture was successful from one needle-exposed deer but not from the tick-exposed deer or dogs. Based on serologic evidence and polymerase chain reaction data, both nymphal and adult ticks transmitted E. chaffeensis to naive deer but not to dogs.

Although more than 320 cases of human ehrlichiosis have been diagnosed in 27 states since 1986, the reservoir host or hosts remain unknown. Since antibodies reactive to Ehrlichia chajyeensis, the etiologic agent of human ehrlichiosis, have been found in white-tailed deer (Odocoileus virginianus), we experimentally evaluated the susceptibilities of four white-tailed deer to infection with E. chaj'eensis and Ehrlichia canis, a

The reservoir hosts of Ehrlichia chaffeensis, etiologic agent of human ehrlichiosis are unknown. Initially, white-tailed deer (WTD) were serologically implicated as possible reservoirs of E. chaffeensis. Subsequent studies showed that WTD were susceptible to infection with E. chaffeensis and that deer-to-deer transmission by a tick vector, Amblyomma americanum, is possible under experimental conditions. To determine if wild WTD were infected with E. chaffeensis, whole blood was collected from 10 deer from Oklahoma and Georgia. All 10 deer had antibodies reactive to E. chaffeensis. Whereas E. chaffeensis was not isolated, restriction enzyme mapping and sequencing of the 16S rDNA gene revealed that a unique Ehrlichia-like agent was present. All 10 deer appeared to be infected with the same agent. We suspect that A. americanum is the vector of this new agent based upon the previously published temporal association between the appearance of E. chaffeensis seropositive WTD and A. americanum. However, the taxonomic and antigenic relationships, geographic distribution, epidemiology, and zoonotic potential of this agent are yet to be determined.

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