Wong S J, Brady G S, Dumler J S. Maryland were found by IFA testing to have antibodies to both the HGE agent and antibodies by immunoblotting. These results suggest that white-tailed deer in diverse geographical regions of the United States are naturally infected with the HGE agent, ticks are known to be vectors for transmission of the HGE agent (19, 21). Transmission from nymphal-stage ticks occurs predominantly during the summer months of May through July, a period which coincides with the seasonal distribution of SJ 172550 the majority of cases of HGE (4). If infected adult ticks feed on large mammals, such as deer, these mammals may serve as sentinels for regions where there is a high risk for transmission (5). Deer participate in the maintenance of the tick life cycle as hosts for adult stages, but SJ 172550 their role as reservoirs is usually controversial. White-tailed deer (species and are a proven reservoir for (16). Recently, Dawson et al. described the presence of novel species 16S rRNA gene sequences in the blood of white-tailed deer with antibodies and interpreted the findings as SJ 172550 evidence of infection with a new uncultured species (6). The presence of a high rate of natural contamination in deer by CD80 such species is problematic when indirect immunofluorescent antibody (IFA) assessments are used, owing to serologic cross-reactivity among tick-transmitted species. Therefore, the use of immunoblots that employ specific HGE agent or antigens can be useful in identifying the infecting species (7, 24). In order to assess whether deer may become naturally infected by the HGE agent or and act as markers of natural transmission or as reservoirs of contamination, we performed IFA assessments and immunoblots on white-tailed deer from northwest Wisconsin and Maryland. MATERIALS AND METHODS Sample collection. Blood was obtained from the peritoneal cavities SJ 172550 of 43 deer shot during the 1994 fall hunting season and from 294 deer during the 1995 fall hunting season in northwestern Wisconsin. The 1994 hunt season deer sera were collected at one checkpoint site in Washburn County, and the 1995 hunt season deer sera were collected in six counties of northwestern Wisconsin, including Barron, Bayfield, Burnett, Douglas, Sawyer, and Washburn Counties, that have a high population density of ticks and reported cases of HGE. The sera were separated from clotted blood and stored frozen at ?20C until used. Sera from 12 southwestern Maryland deer, collected in Charles County in 1992 to 1993, were provided courtesy of Abdu F. Azad, University of Maryland School of Medicine. IFA testing. Serum samples from white-tailed deer were tested for either or HGE agent antibodies and for antibodies with the IFA test (7). MRK or the HGE agent Webster strain cultivated in HL60 cells (11) and (Arkansas stress; thanks to J. Dawson, Centers for Disease Avoidance and Control, Atlanta, Ga.) cultivated in DH82 cells had been utilized as antigens. Quickly, HL60 and DH82 cells which were around 90 to 100% contaminated with either or the HGE agent and (Arkansas stress) as the antigens (2). Uninfected DH82 and HL60 cell lysates were used as adverse control antigens. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblot planning and staining had been performed as previously referred to (2). Alkaline phosphatase-labeled rabbit anti-deer immunoglobulin G, (Kirkegaard and Perry Laboratories) diluted 1:100 in 1% PBSM and 1% regular rabbit serum, was utilized as a second SJ 172550 antibody. Minimal requirements for interpreting antibodies as owned by the group and by immunoblotting had been rings at 44 kDa for the HGE agent Webster stress antigen with 28 to 29 kDa for the Arkansas stress antigen, respectively. The complete localization of the bands was verified by evaluating the 44-kDa antigen recognized having a monoclonal antibody particular for the group 44-kDa antigen (unpublished data) and with monoclonal antibody 1A9 (thanks to.